Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated Transformation
<i>Rosa roxburghii</i> Tratt. is an emerging fruit endemic to China, which has the reputation of being the “King of Vitamin C” because of its abundance of vitamin C. However, it is also a recalcitrant species that imposes severe limitations on the transformation and whole-plant regenerat...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Horticulturae |
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| Online Access: | https://www.mdpi.com/2311-7524/8/11/1079 |
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| author | Lisha Gong Min Lu Huaming An |
| author_facet | Lisha Gong Min Lu Huaming An |
| author_sort | Lisha Gong |
| collection | DOAJ |
| description | <i>Rosa roxburghii</i> Tratt. is an emerging fruit endemic to China, which has the reputation of being the “King of Vitamin C” because of its abundance of vitamin C. However, it is also a recalcitrant species that imposes severe limitations on the transformation and whole-plant regeneration processes, restricting the verification of the functional genes. Therefore, developing a feasible and efficient genetic transformation method for <i>R. roxburghii</i> is an urgent requirement. Herein, K599 with eGFP was used as the <i>Agrobacterium</i> strain to optimize the genetic transformation from four factors: bacterial concentration, seedling age, infection site, and method. First, the original roots of 5-day-old seedlings were excised, and then the slant cuts of the remaining hypocotyls with 0.5 cm length were placed in K599 at an OD600 of 0.4. Subsequently, the explants were planted in a moistened sterile vermiculite after the beveled site was stained with a clump of bacteria. The results showed that the transformation efficiency of this cutting method was almost 28% at 30 days post-inoculation, while the transformation efficiency obtained by injecting 5-day-old seedlings 0.5–1.0 cm away from the primary root with K599 at an OD600 of 0.4 was only about 7%. Taken together, the current findings provide evidence that <i>Agrobacterium</i>-mediated transformation is a simple, fast, and efficient approach for generating composite <i>R. roxburghii</i> plants. Thus, this method has a broad application to analyze the gene functions in <i>R. roxburghii</i> and other related plant species. |
| first_indexed | 2024-03-09T18:18:32Z |
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| institution | Directory Open Access Journal |
| issn | 2311-7524 |
| language | English |
| last_indexed | 2024-03-09T18:18:32Z |
| publishDate | 2022-11-01 |
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| series | Horticulturae |
| spelling | doaj.art-58559895a0ed4fc2a307b929573af83f2023-11-24T08:29:19ZengMDPI AGHorticulturae2311-75242022-11-01811107910.3390/horticulturae8111079Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated TransformationLisha Gong0Min Lu1Huaming An2Engineering Technology Research Centre for <i>Rosa roxburghii</i> of National Forestry and Grassland Administration, Guiyang 550025, ChinaEngineering Technology Research Centre for <i>Rosa roxburghii</i> of National Forestry and Grassland Administration, Guiyang 550025, ChinaEngineering Technology Research Centre for <i>Rosa roxburghii</i> of National Forestry and Grassland Administration, Guiyang 550025, China<i>Rosa roxburghii</i> Tratt. is an emerging fruit endemic to China, which has the reputation of being the “King of Vitamin C” because of its abundance of vitamin C. However, it is also a recalcitrant species that imposes severe limitations on the transformation and whole-plant regeneration processes, restricting the verification of the functional genes. Therefore, developing a feasible and efficient genetic transformation method for <i>R. roxburghii</i> is an urgent requirement. Herein, K599 with eGFP was used as the <i>Agrobacterium</i> strain to optimize the genetic transformation from four factors: bacterial concentration, seedling age, infection site, and method. First, the original roots of 5-day-old seedlings were excised, and then the slant cuts of the remaining hypocotyls with 0.5 cm length were placed in K599 at an OD600 of 0.4. Subsequently, the explants were planted in a moistened sterile vermiculite after the beveled site was stained with a clump of bacteria. The results showed that the transformation efficiency of this cutting method was almost 28% at 30 days post-inoculation, while the transformation efficiency obtained by injecting 5-day-old seedlings 0.5–1.0 cm away from the primary root with K599 at an OD600 of 0.4 was only about 7%. Taken together, the current findings provide evidence that <i>Agrobacterium</i>-mediated transformation is a simple, fast, and efficient approach for generating composite <i>R. roxburghii</i> plants. Thus, this method has a broad application to analyze the gene functions in <i>R. roxburghii</i> and other related plant species.https://www.mdpi.com/2311-7524/8/11/1079<i>Agrobacterium</i><i>Rosa roxburghii</i> Tratt.hairy root transformationK599composite plants |
| spellingShingle | Lisha Gong Min Lu Huaming An Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated Transformation Horticulturae <i>Agrobacterium</i> <i>Rosa roxburghii</i> Tratt. hairy root transformation K599 composite plants |
| title | Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated Transformation |
| title_full | Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated Transformation |
| title_fullStr | Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated Transformation |
| title_full_unstemmed | Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated Transformation |
| title_short | Generation of Composite <i>Rosa roxburghii</i> Plants with Transgenic Roots by <i>Agrobacterium</i>-Mediated Transformation |
| title_sort | generation of composite i rosa roxburghii i plants with transgenic roots by i agrobacterium i mediated transformation |
| topic | <i>Agrobacterium</i> <i>Rosa roxburghii</i> Tratt. hairy root transformation K599 composite plants |
| url | https://www.mdpi.com/2311-7524/8/11/1079 |
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