Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L.
<i>Abelmoschus manihot</i> L. (Jinhuakui, JHK) is widely cultivated for its pharmacological properties owing to its high flavonoid content and is commonly used as a garden landscape plant. However, the absence of an efficient genetic transformation system poses significant challenges for...
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2025-01-01
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| author | Ting Zhang Jiaqi Hou Hongtao Chu Pengyu Guo Qianzi Sang Zhongxu Liu Li Cao |
| author_facet | Ting Zhang Jiaqi Hou Hongtao Chu Pengyu Guo Qianzi Sang Zhongxu Liu Li Cao |
| author_sort | Ting Zhang |
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| description | <i>Abelmoschus manihot</i> L. (Jinhuakui, JHK) is widely cultivated for its pharmacological properties owing to its high flavonoid content and is commonly used as a garden landscape plant. However, the absence of an efficient genetic transformation system poses significant challenges for functional gene studies in this species. Virus-induced gene silencing (VIGS) is a well-established technique for exploring plant gene functions; however, this technique has not been applied to JHK. Here, a tobacco rattle virus (TRV)–VIGS system was successfully developed for the first time in JHK using the gene encoding phytoene desaturase (<i>AmPDS</i>) as a marker gene. This study investigated the impact of various <i>Agrobacterium</i> infection methods on the efficiency of <i>AmPDS</i> silencing. The results demonstrated that administering two injections—the first on the day of complete cotyledon expansion and the second 14 days later—using pTRV1 and pTRV2–<i>AmPDS</i> cultures resuspended to an OD<sub>600</sub> of 1.0 and via the backside of the blade—led to significant photobleaching in the cotyledons 2 days after the second injection. Subsequent analyses revealed a marked reduction in both chlorophyll content and <i>AmPDS</i> expression. These findings suggest that a VIGS system was successfully developed in JHK, thus providing a rapid and effective method for studying gene function in this species and facilitating future research in JHK genetics. |
| format | Article |
| id | doaj-art-72d7c2d5a1204b6b89ebf13b0e0ae8a6 |
| institution | Kabale University |
| issn | 2223-7747 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Plants |
| spelling | doaj-art-72d7c2d5a1204b6b89ebf13b0e0ae8a62025-01-24T13:46:31ZengMDPI AGPlants2223-77472025-01-0114215010.3390/plants14020150Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L.Ting Zhang0Jiaqi Hou1Hongtao Chu2Pengyu Guo3Qianzi Sang4Zhongxu Liu5Li Cao6Agricultural College, Yanbian University, Yanji 133002, ChinaAgricultural College, Yanbian University, Yanji 133002, ChinaAgricultural College, Yanbian University, Yanji 133002, ChinaLaboratory of Molecular Biology of Tomato, Bioengineering College, Chongqing University, Chongqing 400044, ChinaAgricultural College, Yanbian University, Yanji 133002, ChinaAgricultural College, Yanbian University, Yanji 133002, ChinaAgricultural College, Yanbian University, Yanji 133002, China<i>Abelmoschus manihot</i> L. (Jinhuakui, JHK) is widely cultivated for its pharmacological properties owing to its high flavonoid content and is commonly used as a garden landscape plant. However, the absence of an efficient genetic transformation system poses significant challenges for functional gene studies in this species. Virus-induced gene silencing (VIGS) is a well-established technique for exploring plant gene functions; however, this technique has not been applied to JHK. Here, a tobacco rattle virus (TRV)–VIGS system was successfully developed for the first time in JHK using the gene encoding phytoene desaturase (<i>AmPDS</i>) as a marker gene. This study investigated the impact of various <i>Agrobacterium</i> infection methods on the efficiency of <i>AmPDS</i> silencing. The results demonstrated that administering two injections—the first on the day of complete cotyledon expansion and the second 14 days later—using pTRV1 and pTRV2–<i>AmPDS</i> cultures resuspended to an OD<sub>600</sub> of 1.0 and via the backside of the blade—led to significant photobleaching in the cotyledons 2 days after the second injection. Subsequent analyses revealed a marked reduction in both chlorophyll content and <i>AmPDS</i> expression. These findings suggest that a VIGS system was successfully developed in JHK, thus providing a rapid and effective method for studying gene function in this species and facilitating future research in JHK genetics.https://www.mdpi.com/2223-7747/14/2/150<i>Abelmoschus manihot</i> L.virus-induced gene silencingtobacco rattle virus<i>AmPDS</i>back-of-blade injection |
| spellingShingle | Ting Zhang Jiaqi Hou Hongtao Chu Pengyu Guo Qianzi Sang Zhongxu Liu Li Cao Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L. Plants <i>Abelmoschus manihot</i> L. virus-induced gene silencing tobacco rattle virus <i>AmPDS</i> back-of-blade injection |
| title | Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L. |
| title_full | Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L. |
| title_fullStr | Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L. |
| title_full_unstemmed | Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L. |
| title_short | Establishment of a Virus-Induced Gene Silencing System in <i>Abelmoschus manihot</i> L. |
| title_sort | establishment of a virus induced gene silencing system in i abelmoschus manihot i l |
| topic | <i>Abelmoschus manihot</i> L. virus-induced gene silencing tobacco rattle virus <i>AmPDS</i> back-of-blade injection |
| url | https://www.mdpi.com/2223-7747/14/2/150 |
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