摘要
Eucalyptus is very recalcitrant to in vitro culture.In this research, an efficient shoot organogenesis system was developed using 60-day-old plants of Eucalyptus globulus grown in vitro and non-aerated liquid medium to improve shoot proliferation. Cultures were initiated with hypocotyls and leaf segments from plantlets cultivated on semisolid 1/2 MS modified medium supplemented with 4.44 μM 6-Benzyladenine(BA) and 16.1 μM 1-Naphthaleneacetic acid(NAA). Calli were transferred to shoot induction medium, with either 0.5 or 2.7 μM NAA. Shoot multiplication was carried out on 4.44 μM BA + 0.5 μM NAA medium, and semisolid and non-aerated liquid systems were compared for improving shoot proliferation.Rooting of adventitious shoots was evaluated on medium containing NAA or Indole-3-butyric acid-IBA(5 and16 μM). Callogenesis was obtained from both types of explants, although shoot formation was only obtained from leaf-derived calli. Shoot proliferation on 4.44 μM BA+0.5 μM NAA resulted in the most shoots/callus.Non-aerated liquid medium was more efficient in promoting shoot multiplication(53.5 shoots/callus) than was semisolid medium(28.5 shoots/callus). Levels of phenolic compounds were significantly reduced in the shoots cultivated in liquid medium. Efficient rooting(76%) was obtained using 16 μM IBA.
Eucalyptus is very recalcitrant to in vitro culture.In this research, an efficient shoot organogenesis system was developed using 60-day-old plants of Eucalyptus globulus grown in vitro and non-aerated liquid medium to improve shoot proliferation. Cultures were initiated with hypocotyls and leaf segments from plantlets cultivated on semisolid 1/2 MS modified medium supplemented with 4.44 μM 6-Benzyladenine(BA) and 16.1 μM 1-Naphthaleneacetic acid(NAA). Calli were transferred to shoot induction medium, with either 0.5 or 2.7 μM NAA. Shoot multiplication was carried out on 4.44 μM BA + 0.5 μM NAA medium, and semisolid and non-aerated liquid systems were compared for improving shoot proliferation.Rooting of adventitious shoots was evaluated on medium containing NAA or Indole-3-butyric acid-IBA(5 and16 μM). Callogenesis was obtained from both types of explants, although shoot formation was only obtained from leaf-derived calli. Shoot proliferation on 4.44 μM BA+0.5 μM NAA resulted in the most shoots/callus.Non-aerated liquid medium was more efficient in promoting shoot multiplication(53.5 shoots/callus) than was semisolid medium(28.5 shoots/callus). Levels of phenolic compounds were significantly reduced in the shoots cultivated in liquid medium. Efficient rooting(76%) was obtained using 16 μM IBA.
基金
supported by the National Council for Scientific and Technological Development(CNPq)/Brazil,under Grant 477538/2013-4
