<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of oat (<Emphasis Type="Italic">Avena sativa</Emphasis> L.) cultivars via immature embryo and leaf explants |
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Authors: | Sebastian?Gasparis Cezary?Bregier Waclaw?Orczyk Email author" target="_blank">Anna?Nadolska-OrczykEmail author |
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Institution: | (1) Plant Transformation and Cell Engineering Department, Plant Breeding and Acclimatization Institute, Radzikow, 05-870 Blonie, Poland |
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Abstract: | This paper reports on the successful Agrobacterium-mediated transformation of oat, and on some factors influencing this process. In the first step of the experiments, three
cultivars, two types of explant, and three combinations of strain/vectors, which were successfully used for transformation
of other cereals were tested. Transgenic plants were obtained from the immature embryos of cvs. Bajka, Slawko and Akt and
from leaf base explants of cv. Bajka after transformation with A. thumefaciens strain LBA4404(pTOK233). The highest transformation rate (12.3%) was obtained for immature embryos of cv. Bajka. About 79%
of the selected plants proved to be transgenic; however, only 14.3% of the T0 plants and 27.5% of the T1 showed GUS expression. Cell competence of both types of explant differed in terms of their transformation ability and transgene
expression. The next step of the study was to test the suitability for oat transformation of the pGreen binary vector combined
with different selection cassettes: nptII or bar under the nos or 35S promoter. Transgenic plants were selected in combinations transformed with nos::nptII, 35S::nptII and nos::bar. The highest transformation efficiency (5.3%) was obtained for cv. Akt transformed with nos::nptII. A detailed analysis of the T0 plants selected from a given callus line and their progeny revealed that they were the mixture of transgenic, chimeric-transgenic
and non-transgenic individuals. Southern blot analysis of T0 and T1 showed simple integration pattern with the low copy number of the introduced transgenes. |
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Keywords: | Oat Polyploid cereals Agrobacterium Binary vectors Transgene expression |
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