Abstract
Methods were developed for the production of somatic embryos of asparagus (Asparagus officinalis L.) in suspension culture and subsequent conversion to plantlets on solidified medium. Stem-derived callus that was subcultured twice on Murashige and Skoog (MS) medium + 0.54 μM naphthaleneacetic acid (NAA) and 1.4 μM 2-isopentenyladenine (2iP) was used to initiate suspension cultures. Six out of 15 such cell suspensions (MS medium with 54 to 107 μM NAA) had a high embryogenic capacity. These cell suspensions consisted primarily of single elongated cells (about 90% of all single cells), embryogenic cell clusters (2571/ml), and globular translucent embryos (32/ml). The latter converted to plantlets within four weeks on embryo development medium (EDM), which was solidified MS medium containing 0.54 μM NAA and 0.98 μM 2iP. Suspension-derived embryos formed secondary globular embryos at high frequencies (251 to 258/g callus) when placed on EDM with a low carbohydrate (sucrose, glucose or fructose) level (2%). In contrast, EDM with a high carbohydrate level (10%) caused a reduction in the frequency of secondary embryos (30 to 85/g callus), while resulting in the promotion of embryo growth and conversion, 3.6 to 8.5 times higher than 2% carbohydrates. Transfer of globular somatic embryos from cell suspension to EDM with high carbohydrate levels (4 to 10%) for two weeks followed by transfer to EDM with a low carbohydrate level (2%) resulted in a 2 to 4 times higher conversion rate to plantlets than those that remained at the 4 to 10% levels.
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Abbreviations
- ANOVA:
-
analysis of variance
- EDM:
-
embryo development medium
- NAA:
-
naphthaleneacetic acid
- MS:
-
Murashige and Skoog
- RCBD:
-
randomized complete block design
- 2iP:
-
2-isopentenyladenine
- 2,4-d :
-
2,4-dichlorophenoxyacetic acid
- IAA:
-
indoleacetic acid
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Levi, A., Sink, K.C. Asparagus somatic embryos: Production in suspension culture and conversion to plantlets on solidified medium as influenced by carbohydrate regime. Plant Cell Tiss Organ Cult 31, 115–122 (1992). https://doi.org/10.1007/BF00037695
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DOI: https://doi.org/10.1007/BF00037695