Efficient shoot regeneration from direct apical meristem tissue to produce virus-free purple passion fruit plants

Abstract

Purple passion fruit is an important fresh table fruit. At present, the production of passion fruit is decreasing because of the spread of viral diseases throughout the planting area. The aim of this research was to propagate virus-free plants using a tissue culture technique involving the apical meristem of purple passion fruit. Shoot tips were excised to a length of 2 mm and the shoots were regenerated by culturing on Murashige and Skoog medium supplemented with different concentrations of benzyladenine (BA) (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/l) and 1-naphthaleneacetic acid (NAA) (0.2, 0.5, 1.0, 1.5 and 2.0 mg/l). Root formation was promoted using different concentrations of indole-3-butyric acid (IBA) (0.0, 0.2, 0.4, 0.6, 0.8 and 1.2 mg/l). A greater number of shoots were produced with BA concentrations of 1.0 and 1.5 mg/l than with any other BA concentrations tested (less than 1.0 mg/l or greater than 1.5 mg/l). However, when NAA at any concentration was included in the medium, no shoots were produced in culture. The cultures including 1.0 mg/l and 1.5 mg/l BA were then subcultured four times every two weeks. Initially, the tissue cultured in the 1.5 mg/l BA medium grew faster than that cultured in the 1.0 mg/l BA medium. The tissue cultured with 1.5 mg/l BA generated many short shoots, whereas the tissue cultured with 1.0 mg/l BA, generated long shoots that could be subcultured into individual plants. These regenerated shoots were assayed for the presence of the passion fruit woodiness virus using ELISA or a test strip kit; only virus-free shoots were used for further propagation. Root formation was very good in IBA concentrations of 0.4 and 0.6 mg/l. Thus, virus-free plants could be successfully regenerated directly from the apical meristem.