Transfer of amiprophosmethyl resistance from a Nicotiana plumbaginifolia mutant by somatic hybridisation |
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Authors: | A I Yemets O P Kundel’chuk A P Smertenko V G Solodushko V A Rudas Y Y Gleba Y B Blume |
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Institution: | (1) Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Acad. Zabolotny Street 148, Kiev GSP-22, 252650, Ukraine e-mail: alla@cellbio.freenet.viaduk.net, Tel./Fax: +380 44 266 7104, 2961063, UA |
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Abstract: | Transfer of resistance to the phosphorothioamidate herbicide, amiprophosmethyl (APM), from the β-tubulin mutant of Nicotiana plumbaginifolia to the interspecific N. plumbaginifolia (+)
N. sylvestris and to the intertribal N. plumbaginifolia (+) Atropa belladonna somatic hybrids has been demonstrated. Transfer to the recipient species was accomplished by: (1) symmetric hybridisationand
(2) asymmetric hybridisation using γ-irradiation of donor protoplasts. Cytogenetic analysis confirmed the hybrid origin of
the hybrids obtained. It was established that most of them typically inherited no more than three donor chromosomes, although
it was possible to obtain symmetric hybrids in the case of symmetric fusion. Immunofluorescent microscopy analysis has shown
that protoplasts of the mutant, and of the N. plumbaginifolia (+) N. sylvestris and N. plumbaginifolia (+) A. belladonna hybrids, retained the normal structure of interphase microtubule (MT) arrays and mitotic figures after treatment with 5 μM
APM, whereas MTs of protoplasts of the recipients were destroyed under these conditions. It was also shown that hybrid clones
contained an altered β-tubulin isoform originating from the N. plumbaginifolia mutant. The selected hybrid clones were characterised by cross-resistance to trifluralin, a dinitroaniline herbicide with the same
mode of anti-MT action. Some of the somatic hybrids which could flower were fertile. It was established that seeds of some
fertile hybrids were able to germinate in the presence of 5 μM APM. The results obtained thus support the conclusion that
the technique of somatic hybridisation, especially asymmetric fusion, can be used to transfer APM resistance from the N. plumbaginifolia mutant to different (related and remote) plant species of the Solanaceae, including important crops.
Received: 22 December 1997 / Accepted: 27 July 1999 |
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Keywords: | Amiprophosmethyl β -Tubulin Nicotiana plumbaginifolia mutant Somatic hybridisation Transfer of resistance |
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