Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress
Reed canary grass (<i>Phalaris arundinacea</i> L.) is known for its tolerance to drought, heavy metals, and waterlogging, making it a popular choice for forage production and wetland restoration in the Qinghai-Tibet Plateau (QTP). To accurately assess gene expression in reed canary grass...
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MDPI AG
2023-09-01
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| author | Xuejie Jia Yi Xiong Yanli Xiong Daxu Li Qinqin Yu Xiong Lei Minghong You Shiqie Bai Jianbo Zhang Xiao Ma |
| author_facet | Xuejie Jia Yi Xiong Yanli Xiong Daxu Li Qinqin Yu Xiong Lei Minghong You Shiqie Bai Jianbo Zhang Xiao Ma |
| author_sort | Xuejie Jia |
| collection | DOAJ |
| description | Reed canary grass (<i>Phalaris arundinacea</i> L.) is known for its tolerance to drought, heavy metals, and waterlogging, making it a popular choice for forage production and wetland restoration in the Qinghai-Tibet Plateau (QTP). To accurately assess gene expression in reed canary grass under different abiotic stresses, suitable reference genes need to be identified and validated. Thirteen candidate reference gene sequences were selected and screened using RT-qPCR to detect their expression levels in reed canary grass leaves under drought, salt, cadmium, and waterlogging stresses. Four algorithms were used to assess the stability of the expression levels of the candidate reference genes. The most stably expressed genes were <i>UBC</i> and <i>H3</i> under drought Cd, <i>ETF</i> and <i>CYT</i> under salt stress, and <i>ETF</i> and <i>TUB</i> under waterlogging stress. <i>GAPDH</i> was found to be less stable under abiotic stresses. <i>PIP-1</i>, <i>PAL</i>, <i>NAC 90</i>, and <i>WRKY 72A</i> were selected as response genes for quantitative expression assessment under drought, salt, Cd, and waterlogging stresses to confirm the accuracy of the selected stable reference genes. These results provide a theoretical reference for assessing gene expression in reed canary grass under abiotic stresses. |
| first_indexed | 2024-03-10T22:43:37Z |
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| id | doaj.art-9f1c2982e4d34b249015f2e2424905d2 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-10T22:43:37Z |
| publishDate | 2023-09-01 |
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| series | Genes |
| spelling | doaj.art-9f1c2982e4d34b249015f2e2424905d22023-11-19T10:53:45ZengMDPI AGGenes2073-44252023-09-01149179010.3390/genes14091790Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic StressXuejie Jia0Yi Xiong1Yanli Xiong2Daxu Li3Qinqin Yu4Xiong Lei5Minghong You6Shiqie Bai7Jianbo Zhang8Xiao Ma9College of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaSichuan Academy of Grassland Science, Chengdu 610097, ChinaSichuan Academy of Grassland Science, Chengdu 610097, ChinaSichuan Academy of Grassland Science, Chengdu 610097, ChinaSichuan Academy of Grassland Science, Chengdu 610097, ChinaSchool of Life Science and Engineering, Southwest University of Science and Technology, Mianyang 621002, ChinaSichuan Academy of Grassland Science, Chengdu 610097, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaReed canary grass (<i>Phalaris arundinacea</i> L.) is known for its tolerance to drought, heavy metals, and waterlogging, making it a popular choice for forage production and wetland restoration in the Qinghai-Tibet Plateau (QTP). To accurately assess gene expression in reed canary grass under different abiotic stresses, suitable reference genes need to be identified and validated. Thirteen candidate reference gene sequences were selected and screened using RT-qPCR to detect their expression levels in reed canary grass leaves under drought, salt, cadmium, and waterlogging stresses. Four algorithms were used to assess the stability of the expression levels of the candidate reference genes. The most stably expressed genes were <i>UBC</i> and <i>H3</i> under drought Cd, <i>ETF</i> and <i>CYT</i> under salt stress, and <i>ETF</i> and <i>TUB</i> under waterlogging stress. <i>GAPDH</i> was found to be less stable under abiotic stresses. <i>PIP-1</i>, <i>PAL</i>, <i>NAC 90</i>, and <i>WRKY 72A</i> were selected as response genes for quantitative expression assessment under drought, salt, Cd, and waterlogging stresses to confirm the accuracy of the selected stable reference genes. These results provide a theoretical reference for assessing gene expression in reed canary grass under abiotic stresses.https://www.mdpi.com/2073-4425/14/9/1790reference genes<i>Phalaris arundinacea</i> L.abiotic stressesreal-time quantitative PCR |
| spellingShingle | Xuejie Jia Yi Xiong Yanli Xiong Daxu Li Qinqin Yu Xiong Lei Minghong You Shiqie Bai Jianbo Zhang Xiao Ma Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress Genes reference genes <i>Phalaris arundinacea</i> L. abiotic stresses real-time quantitative PCR |
| title | Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress |
| title_full | Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress |
| title_fullStr | Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress |
| title_full_unstemmed | Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress |
| title_short | Identification and Validation of Reference Genes for RT-qPCR Analysis in Reed Canary Grass during Abiotic Stress |
| title_sort | identification and validation of reference genes for rt qpcr analysis in reed canary grass during abiotic stress |
| topic | reference genes <i>Phalaris arundinacea</i> L. abiotic stresses real-time quantitative PCR |
| url | https://www.mdpi.com/2073-4425/14/9/1790 |
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