Maturation and Conversion of Somatic Embryos Derived from Seeds of Olive (Olea europaea L.) cv. Dahbia: Occurrence of Secondary Embryogenesis and Adventitious Bud Formation

Maturation and conversion of somatic embryos are two crucial steps that hamper the development of efficient somatic embryogenesis systems in olive. Herein, a simple and efficient protocol for the maturation and conversion of olive somatic embryos is reported. Globular somatic embryos derived from seeds of cv. Dahbia were cultured on either half-strength olive (OM) or olive cyclic embryogenesis (ECO) media, with and without plant growth regulators (PGRs). The embryos reached the cotyledonary stage in 9 weeks, but those cultured on ECO medium containing 0.1 mg·L−1 6-(dimethylallylamino)purine (2iP), 0.1 mg·L−1 6-benzyladenine (BA) and 0.05 mg·L−1 indole-3-butyric acid (IBA) exhibited the largest sizes, with an average of 4.7 mm. Somatic embryo conversion into plantlets was evaluated using different culture media (half-strength OM or one-third strength Murashige and Skoog (MS)), light conditions (light or dark) and desiccation pretreatments. The highest rate of somatic embryo conversion (45%) was observed under a 16 h photoperiod on half strength OM medium containing 0.1 mg·L−1 gibberellic acid (GA3) and 0.1 mg·L−1 1-naphthalene acetic acid (NAA). The embryos that failed to germinate showed either necrosis, cotyledon greening with no further conversion, adventitious bud formation or secondary embryogenesis. The findings of this study will be beneficial for biotechnological applications in olive.

In Vitro Factors During Ovule Culture Affect Development and Conversion of Immature Peach and Nectarine Embryos

Various in vitro conditions for culture of ovules prior to extraction and culture of immature embryos of peach [Prunus persica (L.) Batsch] and nectarine [Prunus persica (L.) Batsch var. nucipersica Schneid.] were investigated. Culture vessels consisting of test tubes, petri dishes, and polycarbonate jars were tested along with various types of support and nutrient media. Agar support was superior to liquid media with filter paper supports. Agar produced the largest embryos with 90% to 93% being converted into plants compared to liquid with only 1% to 12% embryo conversion. The best ovule orientation and support was with the micropyle down and pushed halfway into an agar-gelled medium. In experiments two and three, test tubes with vertical ovule orientation (micropyle end of ovule pushed into agar) produced larger embryos, the largest plants and the greatest percentage of embryos that converted into plants (60% and 91%). Petri dish treatments were less successful in embryo conversion than test tubes and polycarbonate jars. The addition of activated charcoal (AC) to an agar-gelled medium produced significantly larger embryos with a similar conversion rate. The addition of an agar-gelled medium to culture vessels reduces preparation time compared to filter paper supports, and placing each ovule within a test tube eliminates cross contamination, making immature embryo culture more successful.

Regeneration of Babaco (Carica pentagona) from Ovular Callus

Ovules of babaco [Carica pentagona (Heilborn) Badillo], 23 to 140 days old, were cultured to initiate regenerative callus. Callus developed from the integuments and possibly from the nucellus. Ovules of greater length and age produced more calli on White's medium or medium with half-strength MS salts than on full-strength MS. Ovules >60 days old that were chilled for 24 hours produced significantly more callus than fresh ovules <60 days old. Ovular calli of summer and fall fruits (73 to 90 days old) grown at 23 ± 2C under cool-white fluorescent lamps (16- or 18-hour photoperiod, 12 or 16 μmol·s-1·m-2) developed green areas that subsequently produced nodular structures. Nodular structures produced proembryonal structures that developed into mature somatic embryos when transferred to media containing either GA3 (0.1 mg·liter-1) plus activated charcoal (2.0 g·liter-1) or casein hydrolysate (200 mg·liter-1) plus IAA (0.5 mg·liter-1). Somatic embryos converted into plantlets when transferred to embryo conversion medium. Chemical names used: 1-H -indole-3-acetic acid (IAA); gibberellic acid (GA3).