An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis
Abstract Background Functional genomic studies using genetics approaches of conifers are hampered by the complex and enormous genome, long vegetative growth period, and exertion in genetic transformation. Thus, the research carried out on gene function in Pinus tabuliformis is typically performed by...
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| Format: | Article |
| Language: | English |
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BMC
2020-04-01
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| Series: | Plant Methods |
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| Online Access: | http://link.springer.com/article/10.1186/s13007-020-00594-5 |
| _version_ | 1818151425798045696 |
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| author | Shuangwei Liu Jingjing Ma Hongmei Liu Yingtian Guo Wei Li Shihui Niu |
| author_facet | Shuangwei Liu Jingjing Ma Hongmei Liu Yingtian Guo Wei Li Shihui Niu |
| author_sort | Shuangwei Liu |
| collection | DOAJ |
| description | Abstract Background Functional genomic studies using genetics approaches of conifers are hampered by the complex and enormous genome, long vegetative growth period, and exertion in genetic transformation. Thus, the research carried out on gene function in Pinus tabuliformis is typically performed by heterologous expression based on the model plant Arabidopsis. However, due to the evolutionary and vast diversification from non-flowering (gymnosperms) to flowering (angiosperms) plants, several key differences may alter the underlying genetic concerns and the analysis of variants. Therefore, it is essential to develop an efficient genetic transformation and gene function identification protocol for P. tabuliformis. Results In the present study we established a highly efficient transgene Agrobacterium-mediated transient expression system for P. tabuliformis. Using a β-glucuronidase gene (GUS) as a reporter gene expression, the highest transformation efficiency (70.1%) was obtained by co-cultivation with Agrobacterium strain GV3101 at an optical density at 600 nm of 0.8, with 150 μM acetosyringone for 30 min followed by 3 days in the dark at 23 ± 1 °C. This protocol would be applied to other conifers; GUS staining was observed 24 h post-infection. Conclusions We report a simple, fast, and resilient system for transient Agrobacterium-mediated transformation high-level expression of target genes in P. tabuliformis, which will also improve transformation efficiency in other conifer species. |
| first_indexed | 2024-12-11T13:38:38Z |
| format | Article |
| id | doaj.art-499dff2e6aca44d99cefb790f880e76f |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-11T13:38:38Z |
| publishDate | 2020-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-499dff2e6aca44d99cefb790f880e76f2022-12-22T01:04:54ZengBMCPlant Methods1746-48112020-04-011611910.1186/s13007-020-00594-5An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformisShuangwei Liu0Jingjing Ma1Hongmei Liu2Yingtian Guo3Wei Li4Shihui Niu5Beijing Advanced Innovation Center for Tree Breeding By Molecular Design, National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry UniversityBeijing Advanced Innovation Center for Tree Breeding By Molecular Design, National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry UniversityBeijing Advanced Innovation Center for Tree Breeding By Molecular Design, National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry UniversityBeijing Advanced Innovation Center for Tree Breeding By Molecular Design, National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry UniversityBeijing Advanced Innovation Center for Tree Breeding By Molecular Design, National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry UniversityBeijing Advanced Innovation Center for Tree Breeding By Molecular Design, National Engineering Laboratory for Tree Breeding, College of Biological Sciences and Technology, Beijing Forestry UniversityAbstract Background Functional genomic studies using genetics approaches of conifers are hampered by the complex and enormous genome, long vegetative growth period, and exertion in genetic transformation. Thus, the research carried out on gene function in Pinus tabuliformis is typically performed by heterologous expression based on the model plant Arabidopsis. However, due to the evolutionary and vast diversification from non-flowering (gymnosperms) to flowering (angiosperms) plants, several key differences may alter the underlying genetic concerns and the analysis of variants. Therefore, it is essential to develop an efficient genetic transformation and gene function identification protocol for P. tabuliformis. Results In the present study we established a highly efficient transgene Agrobacterium-mediated transient expression system for P. tabuliformis. Using a β-glucuronidase gene (GUS) as a reporter gene expression, the highest transformation efficiency (70.1%) was obtained by co-cultivation with Agrobacterium strain GV3101 at an optical density at 600 nm of 0.8, with 150 μM acetosyringone for 30 min followed by 3 days in the dark at 23 ± 1 °C. This protocol would be applied to other conifers; GUS staining was observed 24 h post-infection. Conclusions We report a simple, fast, and resilient system for transient Agrobacterium-mediated transformation high-level expression of target genes in P. tabuliformis, which will also improve transformation efficiency in other conifer species.http://link.springer.com/article/10.1186/s13007-020-00594-5Pinus tabuliformis callusEfficient transformation systemAgrobacterium-mediated transformationTransient gene expressionGUS staining |
| spellingShingle | Shuangwei Liu Jingjing Ma Hongmei Liu Yingtian Guo Wei Li Shihui Niu An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis Plant Methods Pinus tabuliformis callus Efficient transformation system Agrobacterium-mediated transformation Transient gene expression GUS staining |
| title | An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis |
| title_full | An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis |
| title_fullStr | An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis |
| title_full_unstemmed | An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis |
| title_short | An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis |
| title_sort | efficient system for agrobacterium mediated transient transformation in pinus tabuliformis |
| topic | Pinus tabuliformis callus Efficient transformation system Agrobacterium-mediated transformation Transient gene expression GUS staining |
| url | http://link.springer.com/article/10.1186/s13007-020-00594-5 |
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