Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco
Phosphorus deficiency highly interferes with plant growth and development. Plants respond to persistent P deficiency by coordinating the expression of genes involved in the alleviation of stress. Promoters of phosphate transporter genes are a great choice for the development of genetically modified...
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MDPI AG
2023-11-01
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| author | Murugan Naveenarani Huskur Kumaraswamy Mahadeva Swamy Sakthivel Surya Krishna Channappa Mahadevaiah Ramanathan Valarmathi Markandan Manickavasagam Muthukrishnan Arun Govindakurup Hemaprabha Chinnaswamy Appunu |
| author_facet | Murugan Naveenarani Huskur Kumaraswamy Mahadeva Swamy Sakthivel Surya Krishna Channappa Mahadevaiah Ramanathan Valarmathi Markandan Manickavasagam Muthukrishnan Arun Govindakurup Hemaprabha Chinnaswamy Appunu |
| author_sort | Murugan Naveenarani |
| collection | DOAJ |
| description | Phosphorus deficiency highly interferes with plant growth and development. Plants respond to persistent P deficiency by coordinating the expression of genes involved in the alleviation of stress. Promoters of phosphate transporter genes are a great choice for the development of genetically modified plants with enhanced phosphate uptake abilities, which improve crop yields in phosphate-deficient soils. In our previous study, the sugarcane phosphate transporter PHT1;2 gene showed a significantly high expression under salinity stress. In this study, the <i>Erianthus arundinaceus EaPHT1;2</i> gene was isolated and characterized using various in silico tools. The deduced 542 amino acid residues have 10 transmembrane domains, with a molecular weight and isoelectric point of 58.9 kDa and 9.80, respectively. They displayed 71–96% similarity with <i>Arabidopsis thaliana</i>, <i>Zea mays</i>, and the <i>Saccharum</i> hybrid. To elucidate the function of the 5′ regulatory region, the 1.1 kb promoter was isolated and validated in tobacco transgenics under Pi stress. The EaPHT1;2 promoter activity was detected using a β-glucuronidase (GUS) assay. The EaPHT1;2 promoter showed 3- to 4.2-fold higher expression than the most widely used CaMV35S promoter. The 5′ deletion analysis with and without 5′ UTRs revealed a small-sized 374 bp fragment with the highest promoter activity among 5′ truncated fragments, which was 2.7 and 4.2 times higher than the well-used CaMV35S promoter under normal and Pi deprivation conditions, respectively. The strong and short promoter of EaPHT1;2 with 374 bp showed significant expression in low-Pi-stress conditions and it could be a valuable source for the development of stress-tolerant transgenic crops. |
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| last_indexed | 2024-03-11T11:23:49Z |
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| spelling | doaj.art-e43c88b8667e44f1b6c89f541756491d2023-11-10T15:10:26ZengMDPI AGPlants2223-77472023-11-011221376010.3390/plants12213760Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic TobaccoMurugan Naveenarani0Huskur Kumaraswamy Mahadeva Swamy1Sakthivel Surya Krishna2Channappa Mahadevaiah3Ramanathan Valarmathi4Markandan Manickavasagam5Muthukrishnan Arun6Govindakurup Hemaprabha7Chinnaswamy Appunu8Division of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore 641007, Tamil Nadu, IndiaDivision of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore 641007, Tamil Nadu, IndiaDivision of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore 641007, Tamil Nadu, IndiaDivision of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore 641007, Tamil Nadu, IndiaDivision of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore 641007, Tamil Nadu, IndiaDepartment of Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, IndiaDepartment of Biotechnology, Bharathiar University, Coimbatore 641046, Tamil Nadu, IndiaDivision of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore 641007, Tamil Nadu, IndiaDivision of Crop Improvement, Indian Council of Agricultural Research-Sugarcane Breeding Institute, Coimbatore 641007, Tamil Nadu, IndiaPhosphorus deficiency highly interferes with plant growth and development. Plants respond to persistent P deficiency by coordinating the expression of genes involved in the alleviation of stress. Promoters of phosphate transporter genes are a great choice for the development of genetically modified plants with enhanced phosphate uptake abilities, which improve crop yields in phosphate-deficient soils. In our previous study, the sugarcane phosphate transporter PHT1;2 gene showed a significantly high expression under salinity stress. In this study, the <i>Erianthus arundinaceus EaPHT1;2</i> gene was isolated and characterized using various in silico tools. The deduced 542 amino acid residues have 10 transmembrane domains, with a molecular weight and isoelectric point of 58.9 kDa and 9.80, respectively. They displayed 71–96% similarity with <i>Arabidopsis thaliana</i>, <i>Zea mays</i>, and the <i>Saccharum</i> hybrid. To elucidate the function of the 5′ regulatory region, the 1.1 kb promoter was isolated and validated in tobacco transgenics under Pi stress. The EaPHT1;2 promoter activity was detected using a β-glucuronidase (GUS) assay. The EaPHT1;2 promoter showed 3- to 4.2-fold higher expression than the most widely used CaMV35S promoter. The 5′ deletion analysis with and without 5′ UTRs revealed a small-sized 374 bp fragment with the highest promoter activity among 5′ truncated fragments, which was 2.7 and 4.2 times higher than the well-used CaMV35S promoter under normal and Pi deprivation conditions, respectively. The strong and short promoter of EaPHT1;2 with 374 bp showed significant expression in low-Pi-stress conditions and it could be a valuable source for the development of stress-tolerant transgenic crops.https://www.mdpi.com/2223-7747/12/21/3760<i>Erianthus arundinaceus</i>high-affinity phosphate transporterPHT12promoter analysistobacco transgenic |
| spellingShingle | Murugan Naveenarani Huskur Kumaraswamy Mahadeva Swamy Sakthivel Surya Krishna Channappa Mahadevaiah Ramanathan Valarmathi Markandan Manickavasagam Muthukrishnan Arun Govindakurup Hemaprabha Chinnaswamy Appunu Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco Plants <i>Erianthus arundinaceus</i> high-affinity phosphate transporter PHT1 2 promoter analysis tobacco transgenic |
| title | Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco |
| title_full | Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco |
| title_fullStr | Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco |
| title_full_unstemmed | Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco |
| title_short | Isolation and Characterization of <i>Erianthus arundinaceus</i> Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco |
| title_sort | isolation and characterization of i erianthus arundinaceus i phosphate transporter 1 pht1 gene promoter and 5 deletion analysis of transcriptional regulation regions under phosphate stress in transgenic tobacco |
| topic | <i>Erianthus arundinaceus</i> high-affinity phosphate transporter PHT1 2 promoter analysis tobacco transgenic |
| url | https://www.mdpi.com/2223-7747/12/21/3760 |
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