Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCR
Oat (<i>Avena sativa</i> L.) is a widely cultivated cereal with high nutritional value and it is grown mainly in temperate regions. The number of studies dealing with gene expression changes in oat continues to increase, and to obtain reliable RT-qPCR results it is essential to establish...
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MDPI AG
2021-06-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/10/7/1272 |
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| author | Judit Tajti Magda Pál Tibor Janda |
| author_facet | Judit Tajti Magda Pál Tibor Janda |
| author_sort | Judit Tajti |
| collection | DOAJ |
| description | Oat (<i>Avena sativa</i> L.) is a widely cultivated cereal with high nutritional value and it is grown mainly in temperate regions. The number of studies dealing with gene expression changes in oat continues to increase, and to obtain reliable RT-qPCR results it is essential to establish and use reference genes with the least possible influence caused by experimental conditions. However, no detailed study has been conducted on reference genes in different tissues of oat under diverse abiotic stress conditions. In our work, nine candidate reference genes (ACT, TUB, CYP, GAPD, UBC, EF1, TBP, ADPR, PGD) were chosen and analysed by four statistical methods (GeNorm, Normfinder, BestKeeper, RefFinder). Samples were taken from two tissues (leaves and roots) of 13-day-old oat plants exposed to five abiotic stresses (drought, salt, heavy metal, low and high temperatures). ADPR was the top-rated reference gene for all samples, while different genes proved to be the most stable depending on tissue type and treatment combinations. TUB and EF1 were most affected by the treatments in general. Validation of reference genes was carried out by PAL expression analysis, which further confirmed their reliability. These results can contribute to reliable gene expression studies for future research in cultivated oat. |
| first_indexed | 2024-03-10T10:10:01Z |
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| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T10:10:01Z |
| publishDate | 2021-06-01 |
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| spelling | doaj.art-b894f530f8d044b694a70bf107efc96e2023-11-22T01:16:33ZengMDPI AGPlants2223-77472021-06-01107127210.3390/plants10071272Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCRJudit Tajti0Magda Pál1Tibor Janda2Department of Plant Physiology, Agricultural Institute, Centre for Agricultural Research, ELKH, H-2462 Martonvásár, HungaryDepartment of Plant Physiology, Agricultural Institute, Centre for Agricultural Research, ELKH, H-2462 Martonvásár, HungaryDepartment of Plant Physiology, Agricultural Institute, Centre for Agricultural Research, ELKH, H-2462 Martonvásár, HungaryOat (<i>Avena sativa</i> L.) is a widely cultivated cereal with high nutritional value and it is grown mainly in temperate regions. The number of studies dealing with gene expression changes in oat continues to increase, and to obtain reliable RT-qPCR results it is essential to establish and use reference genes with the least possible influence caused by experimental conditions. However, no detailed study has been conducted on reference genes in different tissues of oat under diverse abiotic stress conditions. In our work, nine candidate reference genes (ACT, TUB, CYP, GAPD, UBC, EF1, TBP, ADPR, PGD) were chosen and analysed by four statistical methods (GeNorm, Normfinder, BestKeeper, RefFinder). Samples were taken from two tissues (leaves and roots) of 13-day-old oat plants exposed to five abiotic stresses (drought, salt, heavy metal, low and high temperatures). ADPR was the top-rated reference gene for all samples, while different genes proved to be the most stable depending on tissue type and treatment combinations. TUB and EF1 were most affected by the treatments in general. Validation of reference genes was carried out by PAL expression analysis, which further confirmed their reliability. These results can contribute to reliable gene expression studies for future research in cultivated oat.https://www.mdpi.com/2223-7747/10/7/1272reference gene validationRT-qPCRabiotic stressoat |
| spellingShingle | Judit Tajti Magda Pál Tibor Janda Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCR Plants reference gene validation RT-qPCR abiotic stress oat |
| title | Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCR |
| title_full | Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCR |
| title_fullStr | Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCR |
| title_full_unstemmed | Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCR |
| title_short | Validation of Reference Genes for Studying Different Abiotic Stresses in Oat (<i>Avena sativa</i> L.) by RT-qPCR |
| title_sort | validation of reference genes for studying different abiotic stresses in oat i avena sativa i l by rt qpcr |
| topic | reference gene validation RT-qPCR abiotic stress oat |
| url | https://www.mdpi.com/2223-7747/10/7/1272 |
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