Biochemical and Physiological Changes of Callus Growth and t-anethole Essential Oil Production from pimipinella anisum L. under Salinity Stress


This experiment was performed in faculty of Science labs, Babylon University, carried out during 2014-2015 for extraction, and Quantitative of essential oil t-anethole , from seeds , leaves and callus tissue of (pimipinella anisum L.). This study include sterilized anise seeds were germinated in free MS medium and callus induction from leaves (2cm pieces) by using same MS medium supplemented with plant regulator 2,4-Dichlorophenoxiactic acid (2,4-D) at different concentration (1, 2, 3) mg/l (induced phase). Callus tissue induced sub-cultured on MS medium with (BA) benzyl adenine at concentration of (1, 2, 3) mg/l and 2,4-D at concentration of 2 mg/l. Then, identically callus fresh weight sub-cultured on the same MS Medium supplemented with different concentration of NaCl (6, 9, 12 and 15Ds/m), Finally, comparisons study were made between in vitro and in vivo grown plant. Solution of control (t-anethole) used as standard. The content of t-anethole compound was done by using HPLC (high performance liquid chromatography), and compare of the quantity of t-anethole in callus with these content in seeds and leaves of mother plant. The result showed significant increased (P<0.05) of t-anethole production in callus tissue than the content in seeds and leaves of mother plant. Antioxidant enzymes activity like Catalase (CAT), were high in callus under different stresses conditions than in callus grown without stress. However, Proline was higher levels in callus under salt stress than in callus grown without stress