Plant regeneration in vitro, characterization and control of vitrification, and somaclonal variation in onion (Allium cepa L.)
Schloupt, Renee Marie
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https://hdl.handle.net/2142/23768
Description
Title
Plant regeneration in vitro, characterization and control of vitrification, and somaclonal variation in onion (Allium cepa L.)
Author(s)
Schloupt, Renee Marie
Issue Date
1994
Doctoral Committee Chair(s)
Splittstoesser, Walter E.
Department of Study
Agriculture, Plant Culture
Discipline
Agriculture, Plant Culture
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Agriculture, Plant Culture
Language
eng
Abstract
Several systems were tested for the induction of vitrification so that causes could be studied and possible prevention measures developed. High levels of vitrification were obtained when explants were placed on cosmetic puffs suspended in liquid medium or on medium solidified with low agar levels. Vitrification also significantly increased when decreasing concentrations of gelrite were used to solidify the medium. BA concentrations ranging from 0 to 71 uM had no effect on vitrification level. When varying BA, gelrite and agar concentrations were combined in a factorial design, there was no significant difference in vitrification between individual BA and gelling agent combinations.
Scanning electron microscope studies revealed a difference in leaf morphology between vitrified onion leaves, normal micropropagated leaves and leaves from glasshouse-grown plants. Vitrified leaves appeared to have little epicuticular wax. Stomata were raised above the epidermis, round in shape, and many had malformed openings.
Vitrification could not be eliminated from onion tissue cultures. However, adding activated charcoal to the tissue culture medium or placing explants in a small desiccator containing silica gel significantly reduced percentage vitrification over control plants.
Varying concentrations of the fungicides Captan and Benomyl had no significant effect on the level of fungal contamination or plant growth in onion tissue cultures.
There is little problem removing onion plants from tissue culture and acclimating them to glasshouse conditions. Significant differences were found between onion clones for foliage leaf and bulb ring number. However, these measures may be unreliable due to a fungal infection, which inhibited growth. Flow cytometry is a fast, efficient alternative to chromosome counting to determine ploidy levels. Cytometric analysis revealed one chimeric multiploid in clone 117. Statistical analysis of the G1 DNA content of the clones revealed the possibility of chromosomal abnormalities such as breakages and deletions in several of the clones.
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