Enhanced seed germination of Ilex dumosa R. (Aquifoliaceae) through in vitro culture of cut pyrenes

Abstract

An in vitro culture protocol was developed that increased the germination percentage and decreased the lag time to germination for Ilex dumosa R. pyrenes as a tool for replacing the laborious task of embryo rescue technique. This method involves transversely cutting surface-sterilized pyrenes with a scalpel blade, then placing the micropylar one-third end with the rudimentary embryo (0.25 mm long) on solidified (agar 0.65%) quarter-strength salts and vitamins of Murashige and Skoog, 1962 medium with 3% sucrose, and incubating in a growth room at 27 ± 2 8C with a 14-h photoperiod (116 mmolm?2 s?1). Most of the cut pyrenes (greater than 50%) germinated within the first month after inoculation and achieved maximum germination (70%) in 2 months compared with whole pyrenes, which began to germinate 3 months after sowing and required more than 8 months for maximum germination (37%). Moreover, the germination percentage of cut pyrenes was significantly higher than the germination of isolated embryos (34%). Thus, the cut pyrenes culture is a simpler and more effective technique than embryo rescue. Easily, on average, a trained operator is able to culture ~1000 cut pyrenes per day instead of ~100 isolated embryos.Fil: Dolce, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; ArgentinaFil: Mroginski, Luis Amado. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; ArgentinaFil: Rey, Hebe Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentin