An easy and reliable method for establishment and maintenance of tissue cultures of Nicotiana tabacum cv TAPM 26

Abstract

In vitro regeneration of Nicotiana tabacum was efficiently achieved using growth regulators combination supplemented into semi-solid MS medium using leaves from 1-month-old plant as explants. The growth regulators used were NAA for auxin (0.0, 1.0, 2.0, and 3.0 mgL -1 ) and BAP (0.0, 0.5, 1.0, 2.5, and 5.0 mgL -1 ). The effects of growth regulator combination were assessed based on a number of callus formation, shoots formation and fresh callus weight. The maximum number of callus formation was 100 % at five hormone combination, observed at the fourth week after culture. Maximum number of shoots produced per explant was 21.4 shoots at 1.0 mgL -1 BAP + 3.0 mgL -1 NAA, after 6 weeks of culture. The maximum callus fresh weights were obtained at 0.5 mgL -1 BAP + 1.0 mgL -1 NAA after 6 weeks (9.92 g). The best combination for shoots regeneration of Nicotiana tabacum was 1.0 mgL -1 BAP + 3.0 mgL -1 NAA. And the best combination for heaviest callus production was 0.5 mgL -1 BAP + 1.0 mgL -1 NAA.