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A tobacco clone TPD1 (Tobacco Pollen Development), characterized by an extended flowering period, was obtained using a serial agrobacterial transformation of tobacco (Nicotiana tabacum L., cv. Samsun) by constructing a DNA carrying the kanamycin resistance gene inserted into the binary pBin19 vector. However, the characteristics of the vegetative growth of these plants were similar to those of other tobacco clones and the wild type. In the insertion mutant, pollen was 1.5 times smaller than in the wild type and germinated on the stigmata of neither its own clone nor the wild-type plants. Cytochemical investigation of pollen of the TPD1-mutant did not reveal any activity of respiratory enzymes, succinate dehydrogenase and peroxidase, indicating that the pollen was nonviable. Unlike the wild type, the TPD1 mutant exhibited disturbed trophic interactions within the anther tissues, particularly suppressed starch hydrolysis in the anther wall tissues at the stage of rapidly growing microspores. We conclude that the insertion of T-DNA into the TPD1 gene produced structural and metabolic changes during the development of anther tissues in the mutant clone, resulting in pollen sterility.