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Polyacrylamide Gel Electrophoresis for Cultivar Identification in Tobacco¹

Polyacrylamide gel electrophoresis (PAGE) has been demo onstrated as a potential tool for cultivar identification in several crop species; however, information concerning tobacco is generally lacking in the literature. Plants from 10 cultivars representing Maryland and burley air-cured, flue-cured, and cigar tobacco types were grown in the field and in two separate greenhouses. Leaf samples were collected and frozen in liquid N₂ at three maturity dates (32, 40 and 60 days after transplanting). Isozyme separations were performed using PAGE for esterase, catalase, malate dehydrogenase, and peroxidase with the number of bands identified for each enzyme combined over cultivars and treatments being 3, 7, 9, and 11, respectively. Leaf material from the earliest maturity dates exhibited the highest number of isozymes and the greenhouse environments produced fewer bands per enzyme than the field environment. No cultivar differences were observed for esterase; however, cultivar groupings were observed for the three other enzymes investigated. Peroxidase and catalase exhibited the greatest potential for use in cultivar identification. Identification of the 10 cultivars using a single enzyme at a given maturity date was not accomplished; however, it was possible to separate the cultivars using one or more enzymes over environments and maturity dates. The technique does have potential for practical application; however, some additional refinements in procedures appear in order to reduce the time requirements during extraction.

Key Words: Biochemical indicator • Plant breeding • Nicotina tabacum L.

² Graduate assistant, associate, and professor, respectively, Dep. of Agronomy, Univ. of Maryland, College Park, MD 20742.

Received for publication July 30, 1984.