title: Efficient induction of apospory and apogamy in vitro in silver fern (Pityrogramma calomelanos L.).
abstract: Silver fern (Pityrogramma calomelanos L.) is a terrestrial or lithophytic herbaceous fern used for ornamental and medicinal purposes. In its farina it produces the cytotoxic and anticancer compound dihydrochalcone. In vitro induction of apospory and apogamy, and direct field establishment of aposporous gametophytes and subsequent sporophyte development has been accomplished. Half-strength Murashige and Skoog (MS) medium with 3.33 microM N(6)-benzyladenine (BA) and 2.32 microM kinetin (Kn) showed earlier development and produced higher numbers of aposporous gametophytes than half-strength MS basal medium. Crozier explants developed higher numbers (mean value 29.2) of gametophytes, but were slower than frond explants (mean value 23.2). The gametophytes originated from the epidermal hairs progressed from uniseriate filamentous to cordate through bi-, tri- and multiseriate and spatulate stage with the development of antheridia. Reduction in the nutrient and sucrose concentrations in the media favoured apogamy. Sucrose-free 1/10 strength MS medium and agar plates developed a mean of 30.4 and 29.9 sporophytes, respectively in the light. The greenhouse-established gametophytes developed sporophytes. The established sporophytes ex vitro showed 95% survival rate. Apogamous sporophytes and the source plant showed the same chromosome numbers (2n=116). The established protocol accomplishes apogamy and apospory in silver fern, and the aposporous gametophytes can be used for genetic transformation and development of transgenic silver fern.
authors.affiliation: Department of Biotechnology, University of Calicut, Kerala, 673635, India. marttinet@yahoo.com
authors.forename: Kottackal Poulose
authors.initials: KP
authors.lastname: Martin
authors.forename: S
authors.initials: S
authors.lastname: Sini
authors.forename: Chun-Lai
authors.initials: CL
authors.lastname: Zhang
authors.forename: Adrian
authors.initials: A
authors.lastname: Slater
authors.forename: P V
authors.initials: PV
authors.lastname: Madhusoodanan
mesh_terms: D029624:Ferns / Q000187:drug effects / Q000502:physiology*; D005854:Germ Cells / Q000187:drug effects / Q000502:physiology*; D010937:Plant Growth Regulators / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects; D018517:Plant Roots / Q000187:drug effects; D013170:Spores / Q000502:physiology*; D013395:Sucrose / Q000494:pharmacology; D046509:Tissue Culture Techniques
chemical_list: D010937:Plant Growth Regulators; D013395:Sucrose