Transmission genetics of Nicotiana hybrids produced by protoplast fusion |
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Authors: | Yu Yu Gleba I F Kanevsky N N Cherep |
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Institution: | (1) Institute of Botany, Academy of Sciences of the Ukrainian S.S.R., Repina 2, 252601 Kiev, USSR;(2) Present address: DNA Plant Technology Corporation, 2611 Branch Pike, 08077 Cinnaminson, NJ, USA |
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Abstract: | Somatic cell hybrids were produced by fusing protoplasts isolated from callus cells of a tobacco line transformed by Agrobacterium tumefaciens (octopine synthesizing strain B6S3), and mesophyll protoplasts from haploid plants of Nicotiana plumbaginifolia. Hybrids were selected by using differential medium (hormone-independent growth plus greening capacity), or by mechanical
isolation and cloning of individual heterokaryocytes. The analysis of hybrid cell lines included the determination of lysopine
dehydrogenase activity (encoded by the T-region of Agrobacterium tumefaciens plasmid), examination of isozymes of esterase, and study of chromosome number and morphology. All eight cell lines selected
on the screening medium were identified as nuclear hybrids, while only three of the eight evaluated clones obtained by mechanical
isolation and cloning were found to be nuclear hydrids; the rest of them were nuclear segregants of tobacco 1] or N. plumbaginifolia 4] type. These data give independent evidence for the occurrence of non-fusion and segregation of nuclei in fusion products,
that can be revealed only by using nonselective methods for hybrid screening. In this paper we demonstrate the value of microisolation
for the recovery of cytoplasmic hybrids. |
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Keywords: | Nicotiana protoplast fusion hybrids genetic constitution T-DNA |
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