Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal Treatments
Best known as licorice, <i>Glycyrrhiza</i> Linn., a genus of herbaceous perennial legume, has been used as a traditional herbal medicine in Asia and a flavoring agent for tobacco and food industry in Europe and America. Abiotic stresses and hormonal treatments can significantly impact th...
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MDPI AG
2020-10-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/9/11/1441 |
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| author | Yuping Li Xiaoju Liang Xuguo Zhou Zhigeng Wu Ling Yuan Ying Wang Yongqing Li |
| author_facet | Yuping Li Xiaoju Liang Xuguo Zhou Zhigeng Wu Ling Yuan Ying Wang Yongqing Li |
| author_sort | Yuping Li |
| collection | DOAJ |
| description | Best known as licorice, <i>Glycyrrhiza</i> Linn., a genus of herbaceous perennial legume, has been used as a traditional herbal medicine in Asia and a flavoring agent for tobacco and food industry in Europe and America. Abiotic stresses and hormonal treatments can significantly impact the development and metabolism of secondary metabolites in <i>Glycyrrhiza</i>. To better understand the biosynthesis of the trace-amount bioactive compounds, we first screened for the suitable reference genes for quantitative real-time reverse transcription PCR (qRT-PCR) analysis in <i>Glycyrrhiza.</i> The expression profiles of 14 candidate reference genes, including <i>Actin1</i> (<i>ACT</i>)<i>, Clathrin complex AP1</i> (<i>CAC</i>), <i>Cyclophilin</i> (<i>CYP</i>), <i>Heat-shock protein 40</i> (<i>DNAJ</i>), <i>Dehydration responsive element binding gene</i> (<i>DREB</i>), <i>Translation elongation factor1</i> (<i>EF1</i>), <i>Ras related protein</i> (<i>RAN</i>), <i>Translation initiation factor</i> (<i>TIF1</i>), <i>β-Tubulin</i> (<i>TUB</i>), <i>Ubiquitin-conjugating enzyme E2</i> (<i>UBC2</i>), <i>ATP binding-box transpoter 2</i> (<i>ABCC2</i>), <i>COP9 signal compex subunit 3</i> (<i>COPS3</i>), <i>Citrate synthase</i> (<i>CS</i>), and <i>R3H domain protein 2</i> (<i>R3HDM2</i>) from two congeneric species, <i>Glycyrrhiza uralensis</i> F. and <i>Glycyrrhiza inflata</i> B., were examined under abiotic stresses (osmotic and salinity) and hormonal treatments (Abscisic acid (ABA) and methyl jasmonic acid (MeJA)) using a panel of software, including geNorm, NormFinder, BestKeeper, and Delta CT. The overall stability, however, was provided by RefFinder, a comprehensive ranking system integrating inputs from all four algorithms. In <i>G. uralensis</i>, the most stable reference genes under osmotic stress, salt stress, ABA treatment, and MeJA treatment were <i>TIF1</i>, <i>DNAJ</i>, <i>CS</i>, and <i>ABCC2</i> for leaves and <i>DNAJ</i>, <i>DREB</i>, <i>CAC</i>, and <i>CAC</i> for roots, respectively. In comparison, the top ranked genes were <i>TUB</i>, <i>CAC</i>, <i>UBC2</i>, and <i>RAN</i> for leaves and <i>TIF1</i>, <i>ABCC2</i>, <i>CAC</i>, and <i>UBC2</i> for roots, respectively, under stress and hormonal treatments in <i>G. inflata</i>. <i>ACT</i> and <i>TIF1</i>, on the other hand, were the least stable genes under the most experimental conditions in the two congeneric species. Finally, our survey of the reference genes in legume shows that <i>EF</i>, <i>ACT</i>, <i>UBC2</i>, and <i>TUB</i> were the top choices for the abiotic stresses while <i>EF</i>, <i>UBC2</i>, <i>CAC</i>, and <i>ABCC2</i> were recommended for the hormonal treatments in Leguminosae. Our combined results provide reliable normalizers for accurate gene quantifications in <i>Glycyrrhiza</i> species, which will allow us to exploit its medicinal potential in general and antiviral activities in particular. |
| first_indexed | 2024-03-10T15:20:58Z |
| format | Article |
| id | doaj.art-b14bea0199474f91b49d3913f067724c |
| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-03-10T15:20:58Z |
| publishDate | 2020-10-01 |
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| series | Plants |
| spelling | doaj.art-b14bea0199474f91b49d3913f067724c2023-11-20T18:32:17ZengMDPI AGPlants2223-77472020-10-01911144110.3390/plants9111441Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal TreatmentsYuping Li0Xiaoju Liang1Xuguo Zhou2Zhigeng Wu3Ling Yuan4Ying Wang5Yongqing Li6Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement & Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, ChinaKey Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement & Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, ChinaDepartment of Entomology, University of Kentucky, Lexington, KY 40546, USAKey Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement & Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, ChinaKey Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement & Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, ChinaKey Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement & Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, ChinaKey Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement & Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, ChinaBest known as licorice, <i>Glycyrrhiza</i> Linn., a genus of herbaceous perennial legume, has been used as a traditional herbal medicine in Asia and a flavoring agent for tobacco and food industry in Europe and America. Abiotic stresses and hormonal treatments can significantly impact the development and metabolism of secondary metabolites in <i>Glycyrrhiza</i>. To better understand the biosynthesis of the trace-amount bioactive compounds, we first screened for the suitable reference genes for quantitative real-time reverse transcription PCR (qRT-PCR) analysis in <i>Glycyrrhiza.</i> The expression profiles of 14 candidate reference genes, including <i>Actin1</i> (<i>ACT</i>)<i>, Clathrin complex AP1</i> (<i>CAC</i>), <i>Cyclophilin</i> (<i>CYP</i>), <i>Heat-shock protein 40</i> (<i>DNAJ</i>), <i>Dehydration responsive element binding gene</i> (<i>DREB</i>), <i>Translation elongation factor1</i> (<i>EF1</i>), <i>Ras related protein</i> (<i>RAN</i>), <i>Translation initiation factor</i> (<i>TIF1</i>), <i>β-Tubulin</i> (<i>TUB</i>), <i>Ubiquitin-conjugating enzyme E2</i> (<i>UBC2</i>), <i>ATP binding-box transpoter 2</i> (<i>ABCC2</i>), <i>COP9 signal compex subunit 3</i> (<i>COPS3</i>), <i>Citrate synthase</i> (<i>CS</i>), and <i>R3H domain protein 2</i> (<i>R3HDM2</i>) from two congeneric species, <i>Glycyrrhiza uralensis</i> F. and <i>Glycyrrhiza inflata</i> B., were examined under abiotic stresses (osmotic and salinity) and hormonal treatments (Abscisic acid (ABA) and methyl jasmonic acid (MeJA)) using a panel of software, including geNorm, NormFinder, BestKeeper, and Delta CT. The overall stability, however, was provided by RefFinder, a comprehensive ranking system integrating inputs from all four algorithms. In <i>G. uralensis</i>, the most stable reference genes under osmotic stress, salt stress, ABA treatment, and MeJA treatment were <i>TIF1</i>, <i>DNAJ</i>, <i>CS</i>, and <i>ABCC2</i> for leaves and <i>DNAJ</i>, <i>DREB</i>, <i>CAC</i>, and <i>CAC</i> for roots, respectively. In comparison, the top ranked genes were <i>TUB</i>, <i>CAC</i>, <i>UBC2</i>, and <i>RAN</i> for leaves and <i>TIF1</i>, <i>ABCC2</i>, <i>CAC</i>, and <i>UBC2</i> for roots, respectively, under stress and hormonal treatments in <i>G. inflata</i>. <i>ACT</i> and <i>TIF1</i>, on the other hand, were the least stable genes under the most experimental conditions in the two congeneric species. Finally, our survey of the reference genes in legume shows that <i>EF</i>, <i>ACT</i>, <i>UBC2</i>, and <i>TUB</i> were the top choices for the abiotic stresses while <i>EF</i>, <i>UBC2</i>, <i>CAC</i>, and <i>ABCC2</i> were recommended for the hormonal treatments in Leguminosae. Our combined results provide reliable normalizers for accurate gene quantifications in <i>Glycyrrhiza</i> species, which will allow us to exploit its medicinal potential in general and antiviral activities in particular.https://www.mdpi.com/2223-7747/9/11/1441reference geneabiotic stresshormonal treatment<i>Glycyrrhiza</i>medicinal plant |
| spellingShingle | Yuping Li Xiaoju Liang Xuguo Zhou Zhigeng Wu Ling Yuan Ying Wang Yongqing Li Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal Treatments Plants reference gene abiotic stress hormonal treatment <i>Glycyrrhiza</i> medicinal plant |
| title | Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal Treatments |
| title_full | Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal Treatments |
| title_fullStr | Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal Treatments |
| title_full_unstemmed | Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal Treatments |
| title_short | Selection of Reference Genes for qRT-PCR Analysis in Medicinal Plant <i>Glycyrrhiza</i> under Abiotic Stresses and Hormonal Treatments |
| title_sort | selection of reference genes for qrt pcr analysis in medicinal plant i glycyrrhiza i under abiotic stresses and hormonal treatments |
| topic | reference gene abiotic stress hormonal treatment <i>Glycyrrhiza</i> medicinal plant |
| url | https://www.mdpi.com/2223-7747/9/11/1441 |
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