Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther Culture
Production of homozygous lines derived from transgenic plants is one of the important steps for phenotyping and genotyping transgenic progeny. The selection of homozygous plants is a tedious process that can be significantly shortened by androgenesis, cultivation of anthers, or isolated microspores....
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| Format: | Article |
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MDPI AG
2020-07-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/9/7/918 |
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| author | Ludmila Ohnoutková Tomáš Vlčko |
| author_facet | Ludmila Ohnoutková Tomáš Vlčko |
| author_sort | Ludmila Ohnoutková |
| collection | DOAJ |
| description | Production of homozygous lines derived from transgenic plants is one of the important steps for phenotyping and genotyping transgenic progeny. The selection of homozygous plants is a tedious process that can be significantly shortened by androgenesis, cultivation of anthers, or isolated microspores. Doubled haploid (DH) production achieves complete homozygosity in one generation. We obtained transgenic homozygous DH lines from six different transgenic events by using anther culture. Anthers were isolated from T0 transgenic primary regenerants and cultivated in vitro. The ploidy level was determined in green regenerants. At least half of the 2n green plants were transgenic, and their progeny were shown to carry the transgene. The process of dihaploidization did not affect the expression of the transgene. Embryo cultures were used to reduce the time to seed of the next generation. The application of these methods enables rapid evaluation of transgenic lines for gene function studies and trait evaluation. |
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| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T18:21:13Z |
| publishDate | 2020-07-01 |
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| spelling | doaj.art-d5b30e2386634b169d322c2099105c442023-11-20T07:22:16ZengMDPI AGPlants2223-77472020-07-019791810.3390/plants9070918Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther CultureLudmila Ohnoutková0Tomáš Vlčko1Laboratory of Growth Regulators, Palacký University & Institute of Experimental Botany, Czech Academy of Sciences, Šlechtitelů 241/27, 78371 Olomouc, Czech RepublicLaboratory of Growth Regulators, Palacký University & Institute of Experimental Botany, Czech Academy of Sciences, Šlechtitelů 241/27, 78371 Olomouc, Czech RepublicProduction of homozygous lines derived from transgenic plants is one of the important steps for phenotyping and genotyping transgenic progeny. The selection of homozygous plants is a tedious process that can be significantly shortened by androgenesis, cultivation of anthers, or isolated microspores. Doubled haploid (DH) production achieves complete homozygosity in one generation. We obtained transgenic homozygous DH lines from six different transgenic events by using anther culture. Anthers were isolated from T0 transgenic primary regenerants and cultivated in vitro. The ploidy level was determined in green regenerants. At least half of the 2n green plants were transgenic, and their progeny were shown to carry the transgene. The process of dihaploidization did not affect the expression of the transgene. Embryo cultures were used to reduce the time to seed of the next generation. The application of these methods enables rapid evaluation of transgenic lines for gene function studies and trait evaluation.https://www.mdpi.com/2223-7747/9/7/918androgenesis barleydouble haploidembryo culturetransformation |
| spellingShingle | Ludmila Ohnoutková Tomáš Vlčko Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther Culture Plants androgenesis barley double haploid embryo culture transformation |
| title | Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther Culture |
| title_full | Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther Culture |
| title_fullStr | Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther Culture |
| title_full_unstemmed | Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther Culture |
| title_short | Homozygous Transgenic Barley (<i>Hordeum vulgare</i> L.) Plants by Anther Culture |
| title_sort | homozygous transgenic barley i hordeum vulgare i l plants by anther culture |
| topic | androgenesis barley double haploid embryo culture transformation |
| url | https://www.mdpi.com/2223-7747/9/7/918 |
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