Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i>
Modern plant biotechnology often faces the problem of obtaining a stable and powerful vector for gene overexpression. It is known that introns carry different regulatory elements whose effects on transgene expression have been poorly studied. To study the effect of an intron on transgene expression,...
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MDPI AG
2023-04-01
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| Series: | Horticulturae |
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| Online Access: | https://www.mdpi.com/2311-7524/9/4/513 |
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| author | Konstantin V. Kiselev Zlata V. Ogneva Olga A. Aleynova Andrey R. Suprun Alexey A. Ananev Nikolay N. Nityagovsky Alexandra S. Dubrovina |
| author_facet | Konstantin V. Kiselev Zlata V. Ogneva Olga A. Aleynova Andrey R. Suprun Alexey A. Ananev Nikolay N. Nityagovsky Alexandra S. Dubrovina |
| author_sort | Konstantin V. Kiselev |
| collection | DOAJ |
| description | Modern plant biotechnology often faces the problem of obtaining a stable and powerful vector for gene overexpression. It is known that introns carry different regulatory elements whose effects on transgene expression have been poorly studied. To study the effect of an intron on transgene expression, the stilbene synthase 11 (<i>VaSTS11</i>) gene of grapevine <i>Vitis amurensis</i> Rupr. was selected and overexpressed in grapevine callus cell cultures and several plant generations of <i>Arabidopsis thaliana</i> as two forms, intronless <i>VaSTS11c</i> and intron-containing <i>VaSTS11d</i>. The <i>STS</i> genes play an important role in the biosynthesis of stilbenes, valuable plant secondary metabolites. <i>VaSTS11d</i> contained two exons and one intron, while <i>VaSTS11c</i> contained only two exons, which corresponded to the mature transcript. It has been shown that the intron-containing <i>VaSTS11d</i> was better expressed in several generations of transgenic <i>A. thaliana</i> than <i>VaSTS11c</i> and also exhibited a lower level of cytosine methylation. As a result, the content of stilbenes in the <i>VaSTS11d</i>-transgenic plants was much higher than in the <i>VaSTS11c</i>-transgenic plants. Similarly, the best efficiency in increasing the content of stilbenes was also observed in grapevine cell cultures overexpressing the intron-containing <i>VaSTS11d</i> transcript. Thus, the results indicate that an intron sequence with regulatory elements can have a strong positive effect on both transgene expression level and its biological functions in plants and plant cell cultures. |
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| id | doaj.art-27ae04b6d3754de6a886218b634e9124 |
| institution | Directory Open Access Journal |
| issn | 2311-7524 |
| language | English |
| last_indexed | 2024-03-11T04:57:50Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Horticulturae |
| spelling | doaj.art-27ae04b6d3754de6a886218b634e91242023-11-17T19:29:50ZengMDPI AGHorticulturae2311-75242023-04-019451310.3390/horticulturae9040513Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i>Konstantin V. Kiselev0Zlata V. Ogneva1Olga A. Aleynova2Andrey R. Suprun3Alexey A. Ananev4Nikolay N. Nityagovsky5Alexandra S. Dubrovina6Laboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, RussiaLaboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, RussiaLaboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, RussiaLaboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, RussiaLaboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, RussiaLaboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, RussiaLaboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, RussiaModern plant biotechnology often faces the problem of obtaining a stable and powerful vector for gene overexpression. It is known that introns carry different regulatory elements whose effects on transgene expression have been poorly studied. To study the effect of an intron on transgene expression, the stilbene synthase 11 (<i>VaSTS11</i>) gene of grapevine <i>Vitis amurensis</i> Rupr. was selected and overexpressed in grapevine callus cell cultures and several plant generations of <i>Arabidopsis thaliana</i> as two forms, intronless <i>VaSTS11c</i> and intron-containing <i>VaSTS11d</i>. The <i>STS</i> genes play an important role in the biosynthesis of stilbenes, valuable plant secondary metabolites. <i>VaSTS11d</i> contained two exons and one intron, while <i>VaSTS11c</i> contained only two exons, which corresponded to the mature transcript. It has been shown that the intron-containing <i>VaSTS11d</i> was better expressed in several generations of transgenic <i>A. thaliana</i> than <i>VaSTS11c</i> and also exhibited a lower level of cytosine methylation. As a result, the content of stilbenes in the <i>VaSTS11d</i>-transgenic plants was much higher than in the <i>VaSTS11c</i>-transgenic plants. Similarly, the best efficiency in increasing the content of stilbenes was also observed in grapevine cell cultures overexpressing the intron-containing <i>VaSTS11d</i> transcript. Thus, the results indicate that an intron sequence with regulatory elements can have a strong positive effect on both transgene expression level and its biological functions in plants and plant cell cultures.https://www.mdpi.com/2311-7524/9/4/513piceidplant cell culturesresveratroltranscription factorstransgenic cellsvitis |
| spellingShingle | Konstantin V. Kiselev Zlata V. Ogneva Olga A. Aleynova Andrey R. Suprun Alexey A. Ananev Nikolay N. Nityagovsky Alexandra S. Dubrovina Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i> Horticulturae piceid plant cell cultures resveratrol transcription factors transgenic cells vitis |
| title | Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i> |
| title_full | Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i> |
| title_fullStr | Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i> |
| title_full_unstemmed | Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i> |
| title_short | Influence of the 135 bp Intron on Stilbene Synthase <i>VaSTS11</i> Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of <i>Arabidopsis thaliana</i> |
| title_sort | influence of the 135 bp intron on stilbene synthase i vasts11 i transgene expression in cell cultures of grapevine and different plant generations of i arabidopsis thaliana i |
| topic | piceid plant cell cultures resveratrol transcription factors transgenic cells vitis |
| url | https://www.mdpi.com/2311-7524/9/4/513 |
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