Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality Conditions
Selecting an appropriate reference gene is of crucial importance for improving the accuracy of qRT-PCR analyses. In this study, strawberry (<i>Fragaria ananassa</i>) seedlings were subjected to different environmental conditions including heat, cold, drought, salt, white-light, blue-ligh...
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MDPI AG
2021-11-01
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| Series: | Horticulturae |
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| Online Access: | https://www.mdpi.com/2311-7524/7/11/452 |
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| author | Yuntian Ye Yang Lu Guangyi Wang Yongqiang Liu Yunting Zhang Haoru Tang |
| author_facet | Yuntian Ye Yang Lu Guangyi Wang Yongqiang Liu Yunting Zhang Haoru Tang |
| author_sort | Yuntian Ye |
| collection | DOAJ |
| description | Selecting an appropriate reference gene is of crucial importance for improving the accuracy of qRT-PCR analyses. In this study, strawberry (<i>Fragaria ananassa</i>) seedlings were subjected to different environmental conditions including heat, cold, drought, salt, white-light, blue-light, and red-light treatments. The expression levels of seven candidate reference genes, including <i>Fa18S</i>, <i>FaGAPDH</i>, <i>FaPIRUV</i>, <i>FaDBP</i>, <i>FaHISTH4</i>, <i>FaACTIN1</i>, and <i>FaACTIN2</i>, in the strawberry leaves were measured by qRT-PCR. Then, four programs (geNorm, NormFinder, BestKeeper, and RefFinder) were employed as tools to evaluate the expression stability of the candidate reference genes. The results showed that the expression stability of the reference genes varied under different conditions. For the cold stress and white-light treatments, <i>FaACTIN2</i> was evaluated to be the most stable reference gene. <i>FaGAPDH</i> should be used as the reference gene under salt-stress condition and red-light treatment. For the data normalization under drought-stress treatment, <i>FaDBP</i> is the recommended reference gene with the highest expression stability. <i>FaHISTH4</i> was observed to be the best reference gene for data normalization under heat stress and blue-light treatment. This work provides information on selecting reference genes for accurate gene expression analyses of target genes in strawberry leaves under various abiotic stress and light-quality conditions. |
| first_indexed | 2024-03-10T05:28:02Z |
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| id | doaj.art-e0847a8d5fc34697b2a3cad25da9f119 |
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| issn | 2311-7524 |
| language | English |
| last_indexed | 2024-03-10T05:28:02Z |
| publishDate | 2021-11-01 |
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| series | Horticulturae |
| spelling | doaj.art-e0847a8d5fc34697b2a3cad25da9f1192023-11-22T23:33:53ZengMDPI AGHorticulturae2311-75242021-11-0171145210.3390/horticulturae7110452Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality ConditionsYuntian Ye0Yang Lu1Guangyi Wang2Yongqiang Liu3Yunting Zhang4Haoru Tang5College of Horticulture, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Horticulture, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Horticulture, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Horticulture, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Horticulture, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Horticulture, Sichuan Agricultural University, Chengdu 611130, ChinaSelecting an appropriate reference gene is of crucial importance for improving the accuracy of qRT-PCR analyses. In this study, strawberry (<i>Fragaria ananassa</i>) seedlings were subjected to different environmental conditions including heat, cold, drought, salt, white-light, blue-light, and red-light treatments. The expression levels of seven candidate reference genes, including <i>Fa18S</i>, <i>FaGAPDH</i>, <i>FaPIRUV</i>, <i>FaDBP</i>, <i>FaHISTH4</i>, <i>FaACTIN1</i>, and <i>FaACTIN2</i>, in the strawberry leaves were measured by qRT-PCR. Then, four programs (geNorm, NormFinder, BestKeeper, and RefFinder) were employed as tools to evaluate the expression stability of the candidate reference genes. The results showed that the expression stability of the reference genes varied under different conditions. For the cold stress and white-light treatments, <i>FaACTIN2</i> was evaluated to be the most stable reference gene. <i>FaGAPDH</i> should be used as the reference gene under salt-stress condition and red-light treatment. For the data normalization under drought-stress treatment, <i>FaDBP</i> is the recommended reference gene with the highest expression stability. <i>FaHISTH4</i> was observed to be the best reference gene for data normalization under heat stress and blue-light treatment. This work provides information on selecting reference genes for accurate gene expression analyses of target genes in strawberry leaves under various abiotic stress and light-quality conditions.https://www.mdpi.com/2311-7524/7/11/452strawberryreference genesqRT-PCR analysisexpression stability |
| spellingShingle | Yuntian Ye Yang Lu Guangyi Wang Yongqiang Liu Yunting Zhang Haoru Tang Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality Conditions Horticulturae strawberry reference genes qRT-PCR analysis expression stability |
| title | Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality Conditions |
| title_full | Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality Conditions |
| title_fullStr | Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality Conditions |
| title_full_unstemmed | Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality Conditions |
| title_short | Stable Reference Gene Selection for qRT-PCR Normalization in Strawberry (<i>Fragaria × ananassa</i>) Leaves under Different Stress and Light-Quality Conditions |
| title_sort | stable reference gene selection for qrt pcr normalization in strawberry i fragaria ananassa i leaves under different stress and light quality conditions |
| topic | strawberry reference genes qRT-PCR analysis expression stability |
| url | https://www.mdpi.com/2311-7524/7/11/452 |
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