Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to Phytohormones
The rubber elongation factor (REF) is the most abundant protein in the latex of <i>Hevea brasiliensis</i>, which is closely related to natural rubber biosynthesis. In order to gain a deeper understanding of the transcriptional regulation mechanism of <i>HbREF1</i>, a 1758 bp...
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2024-02-01
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| author | Lin-Tao Chen Dong Guo Jia-Hong Zhu Ying Wang Hui-Liang Li Feng An Yan-Qiong Tang Shi-Qing Peng |
| author_facet | Lin-Tao Chen Dong Guo Jia-Hong Zhu Ying Wang Hui-Liang Li Feng An Yan-Qiong Tang Shi-Qing Peng |
| author_sort | Lin-Tao Chen |
| collection | DOAJ |
| description | The rubber elongation factor (REF) is the most abundant protein in the latex of <i>Hevea brasiliensis</i>, which is closely related to natural rubber biosynthesis. In order to gain a deeper understanding of the transcriptional regulation mechanism of <i>HbREF1</i>, a 1758 bp genomic DNA fragment of the <i>HbREF1</i> promoter was isolated. Promoter sequence analysis revealed several transcription factor binding sites in the <i>HbREF1</i> promoter, such as bZIP, bHLH, EIL, AP2/ERF, MYB, and Trihelix. To assess the promoter activity, a series of <i>HbREF1</i> promoter deletion derivatives were created and fused with firefly luciferase (LUC). The LUC image demonstrated that all of the <i>HbREF1</i> promoters exhibited transcriptional activity. Furthermore, the assay revealed the presence of multiple regulatory elements within the promoter region that negatively regulate the transcriptional activity. Subsequent analysis of the transcriptional activity following treatment with phytohormones identified an ABA-responsive element located between −583 bp and −200 bp, an ET-responsive element between −718 bp and −583 bp, a JA-responsive element between −1758 bp and −1300 bp, and a SA-responsive element between −1300 bp and −718 bp. These results were largely consistent with the predictions of cis-acting elements. This study has established significant groundwork for future investigations into the regulatory mechanism of <i>HbREF1</i>. |
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| institution | Directory Open Access Journal |
| issn | 1999-4907 |
| language | English |
| last_indexed | 2024-03-07T22:32:14Z |
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| spelling | doaj.art-06885e6f9ae64469b92c50d17af650d12024-02-23T15:16:48ZengMDPI AGForests1999-49072024-02-0115227610.3390/f15020276Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to PhytohormonesLin-Tao Chen0Dong Guo1Jia-Hong Zhu2Ying Wang3Hui-Liang Li4Feng An5Yan-Qiong Tang6Shi-Qing Peng7School of Life and Health, Hainan University, Haikou 570228, ChinaKey Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture and Rural Affairs, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, ChinaKey Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture and Rural Affairs, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, ChinaKey Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture and Rural Affairs, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, ChinaKey Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture and Rural Affairs, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, ChinaRubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, ChinaSchool of Life and Health, Hainan University, Haikou 570228, ChinaSchool of Life and Health, Hainan University, Haikou 570228, ChinaThe rubber elongation factor (REF) is the most abundant protein in the latex of <i>Hevea brasiliensis</i>, which is closely related to natural rubber biosynthesis. In order to gain a deeper understanding of the transcriptional regulation mechanism of <i>HbREF1</i>, a 1758 bp genomic DNA fragment of the <i>HbREF1</i> promoter was isolated. Promoter sequence analysis revealed several transcription factor binding sites in the <i>HbREF1</i> promoter, such as bZIP, bHLH, EIL, AP2/ERF, MYB, and Trihelix. To assess the promoter activity, a series of <i>HbREF1</i> promoter deletion derivatives were created and fused with firefly luciferase (LUC). The LUC image demonstrated that all of the <i>HbREF1</i> promoters exhibited transcriptional activity. Furthermore, the assay revealed the presence of multiple regulatory elements within the promoter region that negatively regulate the transcriptional activity. Subsequent analysis of the transcriptional activity following treatment with phytohormones identified an ABA-responsive element located between −583 bp and −200 bp, an ET-responsive element between −718 bp and −583 bp, a JA-responsive element between −1758 bp and −1300 bp, and a SA-responsive element between −1300 bp and −718 bp. These results were largely consistent with the predictions of cis-acting elements. This study has established significant groundwork for future investigations into the regulatory mechanism of <i>HbREF1</i>.https://www.mdpi.com/1999-4907/15/2/276<i>Hevea brasiliensis</i>rubber elongation factorpromotertranscriptional regulationphytohormone |
| spellingShingle | Lin-Tao Chen Dong Guo Jia-Hong Zhu Ying Wang Hui-Liang Li Feng An Yan-Qiong Tang Shi-Qing Peng Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to Phytohormones Forests <i>Hevea brasiliensis</i> rubber elongation factor promoter transcriptional regulation phytohormone |
| title | Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to Phytohormones |
| title_full | Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to Phytohormones |
| title_fullStr | Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to Phytohormones |
| title_full_unstemmed | Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to Phytohormones |
| title_short | Functional Analysis of the <i>HbREF1</i> Promoter from <i>Hevea brasiliensis</i> and Its Response to Phytohormones |
| title_sort | functional analysis of the i hbref1 i promoter from i hevea brasiliensis i and its response to phytohormones |
| topic | <i>Hevea brasiliensis</i> rubber elongation factor promoter transcriptional regulation phytohormone |
| url | https://www.mdpi.com/1999-4907/15/2/276 |
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