Efficient induction of apospory and apogamy in vitro in silver fern (Pityrogramma calomelanos L.).
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.
Martin
Kottackal Poulose
KP
Department of Biotechnology, University of Calicut, Kerala, 673635, India. marttinet@yahoo.com
Sini
S
S
Zhang
Chun-Lai
CL
Slater
Adrian
A
Madhusoodanan
P V
PV
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
D010937:Plant Growth Regulators; D013395:Sucrose