Abstract
This study was aimed to optimize the organogenesis in rough lemon (Citrus jambhiri Lush), an important citrus rootstock of India. Organogenesis was induced in epicotyl segments of rough lemon seedlings. Three important factors influencing organogenesis in vitro viz. hormone combination, cut modes and photoperiod were evaluated. Etiolation of seedlings had a positive effect on bud formation and higher number of buds per explant was obtained from etiolated seedlings. Among the cut modes, transverse cut performed better for its effect on the number of regenerated buds and shoots per explant. The best response regarding the number of adventitious buds formed per explant was observed with BAP at the concentration of 2.0 mg/l along with malt extract (500 ppm) and additional dose of 25 g/l sucrose. Key words: Citrus, epicotyl, hypocotyl, etiolation, transverse cut, longitudinal cut, photoperiod.
Highlights
Improvement of citrus by conventional method is hampered by polyembryony, sexual incompatibility and male or female sterility (Grosser and Gmitter, 2005)
This study was aimed to optimize the organogenesis in rough lemon (Citrus jambhiri Lush), an important citrus rootstock of India
Organogenesis was induced in epicotyl segments of rough lemon seedlings
Summary
Improvement of citrus by conventional method is hampered by polyembryony, sexual incompatibility and male or female sterility (Grosser and Gmitter, 2005). Plant improvement via genetic transformation is an alternative to release new cultivars. For developing a routine genetic transformation protocol, an efficient system for in vitro regeneration is required. Photoperiodic factors, cut modes, hormonal concentrations and additives may affect in vitro citrus shoot regeneration. Epicotyl segments excised from seedlings germinated in the dark for 3 - 6 weeks (Luth and Moore, 1999; Yang et al, 2000) and transferred to a 16 h photoperiod that varied from 1 to 3 weeks (Cervera et al, 1998; Pena et al, 1995) improved the transformation efficiency. The effect of auxin on shoot regeneration was rarely concerned, though the main hormone effect on bud formation was Abbreviations: BAP, Benzyl amino purine; NAA, Napthaline acetic acid; IBA, Indoline butyric acid
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