Stable transformation of barley callus using biolistic® particle bombardment and the phosphinothricin acetyltransferase (bar) gene |
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Authors: | Carol M Stiff Andrzej Kilian Huaping Zhou David A Kudrna Andris Kleinhofs |
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Institution: | (1) Department of Biology and Research Center for Cell Differentiation, Seoul National University, 151-742 Seoul, Korea |
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Abstract: | We have previously studied chromosomal and morphological variation in protoplast cultures of diploid petunia (Petunia hybrida) plants. We found that 85% of the regenerants were tetraploid (2n=4x=28). These plants flowered and set seeds.In the present study, the cytological stability of plants regenerated from leaf mesophyll protoplast cultures derived from the progeny of the self-fertile tetraploid plants was assessed on the basis of mitotic analysis, morphological characters, and protein patterns. When we analyzed the root tip chromosomes of 117 regenerants derived from 39 protoclone calluses, all of the regenerants tested retained the parental chromosome number of 2n=4x=28. One hundred regenerants were further analyzed and displayed normal vegetative morphology and retained the floral characteristics of the seed-derived plants from which they were derived. No significant variations in any character were observed among regenerants. When leaf protein patterns from four regenerated tetraploid protoclones were analyzed by two-dimensional polyacrylamide gel electrophoresis and compared with those of seed-derived plants, the protein patterns exhibited great similarity.The data suggest that tetraploidization of petunia plant increases cytological stability during further in vitro cultures and may play an important role in the genetic stability of regenerant populations.Abbreviations BA
benzylaminopurine
- IAA
indole-3-acetic acid
- IEF
isoelectric-focusing
- PVP
polyvinylpyrrolidone
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis |
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Keywords: | cytological stability electrophoresis mesophyll protoplast Petunia hybrida tetraploid |
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