Influence of Vessel Type, Physical State of Medium and Temporary Immersion on the Micropropagation of Three Rhodophiala species

Rhodophiala C. Presl (Amaryllidaceae) is a genus of attractive flowering geophytes native to South America. They have ornamental value, but most species are not well- known and have conservation problems. The objective of this study was to optimize a micropropagation process to support the use and preservation of three Chilean native species, R. montana (Phil.) Traub, R. splendens (Rengifo) Traub., and R. ananuca (Phil.) Traub. The research evaluated the feasibility of implementing liquid medium culture and assessed the influence of different tissue culture systems on the shoot production and biomass increment of small bulbs. Three experiments were carried out. The first one determined the influences of flask size and volume of media; the second compared liquid and solid media, and in the third experiment, a temporary immersion system (TIS), and conventional culture in static liquid, shaken liquid and gelled Murashige and Skoog (MS) media were compared. By using larger (350 mL) flasks with higher (50 mL) media volume, 100% more fresh weight of microbulb was obtained that treatment with smaller flasks (45 mL) and media volume (10 mL). In gelled medium, hyperhydricity affected only 5% of explants, while in liquid medium was 16-40%. Survival to acclimatization reached 87-94% for plants from gelled medium; from liquid medium only 38-69%. TIS yielded higher propagation rate (1.9 shoots in 30 d) compared with shaken liquid medium (1.0) (P < 0.05) in R. ananuca only. Current procedures are appropriate for the support of ex situ conservation and germplasm bank establishment.

[1]  J. Adelberg SIVB 2003 Congress Symposium Proceeding: Plant Growth and Sugar Utilization in an Agitated, Thin-Film Liquid System for Micropropagation , 2004, In Vitro Cellular & Developmental Biology - Plant.

[2]  T. Winkelmann,et al.  In vitro propagation of Hippeastrum × chmielii Chm. – influence of flurprimidol and the culture in solid or liquid medium and in temporary immersion systems , 2005, Plant Cell, Tissue and Organ Culture.

[3]  B. Ruffoni,et al.  The Temporary Immersion System (T.I.S.) for the Improvement of Micropropagation of Ornamental Plants , 2005 .

[4]  M. Bridgen,et al.  Techniques for the In Vitro Propagation of Rhodophiala and Leucocoryne spp. , 2005 .

[5]  C. Wawrosch,et al.  Shoot regeneration from nodules of Charybdis sp.: a comparison of semisolid, liquid and temporary immersion culture systems , 2005, Plant Cell, Tissue and Organ Culture.

[6]  Xueyuan Li,et al.  Optimisation of growing conditions for the apple rootstock M26 grown in RITA containers using temporary immersion principle , 2005, Plant Cell, Tissue and Organ Culture.

[7]  Meira Ziv,et al.  Simple bioreactors for mass propagation of plants , 2005, Plant Cell, Tissue and Organ Culture.

[8]  H. Etienne,et al.  Temporary immersion systems in plant micropropagation , 2002, Plant Cell, Tissue and Organ Culture.

[9]  C. Codina,et al.  In vitro production of bulblets of Cyrtanthus loddigesianus and Cyrtanthus speciosus , 2003 .

[10]  M. A. Smith,et al.  Vessel type, closure, and explant orientation influence in vitro performance of five woody species , 1990 .

[11]  T. Kozai,et al.  Fundamental Studies on Environments in Plant Tissue Culture Vessels , 1986 .

[12]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .