Direct induction of protocorm-like bodies from shoot tips, plantlet formation, and clonal fidelity analysis in Anthurium andreanum cv. CanCan

Abstract

Protocorm-like bodies (PLBs) were induced directly at high frequency from wounded surface of Anthurium andreanum cv. CanCan shoot tip-ends, used as explants. In order to obtain PLB directly, the influence of different types and concentrations of cytokinins were evaluated. Amid the cytokinins, N 6 -(D 2 -isopentenyl)-adenine (2-iP) at a concentration of 15 lM was most effective in inducing PLB whereby *98 (97.8) % of explants induced PLB with an average of 120 PLBs per shoot tip within 50 days of culture. Stereomicroscopic observation meticulously revealed the sequential changes from initiation to maturation of PLB gradually forming shoot apical meristem, shoot primordia and leaf primordia. Mature PLBs showed significant shoot proliferation (98.4 %) in media containing 10 lM 6-furfurylaminopurine forming 17 shoots per PLB within 30 days. The inclusion of activated charcoal (AC) in media containing auxin had promotive effect on rooting whereby 5 lM indole-3-butyric acid plus 500 lM AC resulted in highest number and length of roots. Successfully acclimatized plants, subjected to random amplified polymorphic DNA assessment for genetic fidelity, did not show any variation. Thus, this complete study has successfully outlined a rapid, high frequency direct induction of PLB of Anthurium from shoot tips inclusive of shoot proliferation, rooting and acclimatization.

Direct somatic embryogenesis and genetic homogeneity assessment of regenerated plants of Anthurium andraeanum Linden cv. Fantasia

“…A RAPD-based homogeneity analysis has been successfully used in Zingiber officinales (Rout et al 1998), Camellia spp. (Borchetia et al 2009), A. andraeanum (Gantait et al 2012), and Musa sp. (Punyarani et al 2013).…”

“…Among the several DNAbased molecular markers, RAPD has been used to demonstrate the genetic fidelity of in vitro propagated plants of A. andraeanum (Gantait et al 2012). However, no report using RAPD has been published for a genetic uniformity study of direct SE-derived plants of A. andraeanum.…”

Anthurium in vitro: A review

“…There are different possible sources of explants in Anthurium: spadices, spathes or seeds (Wang et al, 1998;Vargas et al, 2004;Alves dos Santos et al, 2005;Jahan et al, 2009;Maira et al, 2010;Winarto et al, 2011a;Ancy et al, 2012), leaves (Eapen and Rao, 1985;Geier, 1986;Teng, 1997;Joseph et al, 2003;Lan et al, 2003b;Martin et al, 2003;Puchooa, 2005;Nhut et al, 2006;Viégas et al, 2007;Bejoy et al, 2008;Atak and Ç elik, 2009;Seah, 2009;Islam et al, 2010;Kumari et al, 2011;Reddy et al, 2011;Farsi et al, 2012), petioles and pedicels (Eapen and Rao, 1985;Raad et al, 2012a), and shoot tips (Gantait et al, 2008(Gantait et al, , 2012Liendo and Mogollón, 2009). Different methods were applied to surface sterilize explants, but these depended on the explant type and Anthurium species.…”

“…Kuehnle et al (1992) had earlier used tree-fern fiber to acclimatize in vitro propagated plants. Gantait et al (2012) acclimatized plants that had been derived from shoot-tip-induced protocorm-like bodies (PLBs) (a possible misnomer) by first washing individual plantlets in water, then by dipping the basal section in a 0.2% solution of Bavistin ® DF, a fungicide, for 2 min. Roots were trimmed and plantlets were transferred into earthen pots containing a mixture of autoclaved sand and soil (2:1, v/v) and covered with plastic lids and watered intermittently.…”

“…An observation of the ultrastructural features in the Kuehnle et al (1992), Chen et al (1997) and Gantait et al (2012) papers indicates that somatic embryos and PLBs (a term used only by the last two papers) are one and the same organ. However, PLBs are generally reserved exclusively for orchids and thus the use of the term PLB might not be appropriate for Anthurium.…”