Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental Conditions
<i>Ginkgo biloba</i>, a deciduous tree species in the Ginkgo family, has a long history of cultivation in China and is widely used in garden landscapes, medicine, food, and health products. However, few reports have focused on the systematic selection of optimal reference genes based on...
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MDPI AG
2020-11-01
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| Series: | Forests |
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| Online Access: | https://www.mdpi.com/1999-4907/11/11/1217 |
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| author | Tingting Zhou Xiaoming Yang Fangfang Fu Guibin Wang Fuliang Cao |
| author_facet | Tingting Zhou Xiaoming Yang Fangfang Fu Guibin Wang Fuliang Cao |
| author_sort | Tingting Zhou |
| collection | DOAJ |
| description | <i>Ginkgo biloba</i>, a deciduous tree species in the Ginkgo family, has a long history of cultivation in China and is widely used in garden landscapes, medicine, food, and health products. However, few reports have focused on the systematic selection of optimal reference genes based on transcriptomic data in <i>G. biloba</i>. The purpose of our research was to select an internal reference gene suitable for different experimental conditions from thirteen candidate reference genes by the delta cycle threshold (ΔCt) method, geNorm, BestKeeper, NormFinder, and RefFinder programs. The reference genes were used for gene expression analyses of <i>Ginkgo biloba</i>. These results showed that elongation factor 1(<i>EF1</i>) and ubiquitin (<i>UBI)</i> were the best choices for samples of different ginkgo genotypes. The expression of <i>UBI</i> and <i>HAS28</i> presented the most stable at different developmental stages of ginkgo, and <i>EIF3I</i> and <i>RPII</i> were considered as suitable reference genes in different tissues of ginkgo. For methyl jasmonate (MeJA) treatment, <i>ACA</i> and <i>ACT</i> were identified as the optimal reference genes. For cold stress treatment, <i>RPII</i> and <i>EIF4E</i> were chosen for the gene expression normalizations. <i>HAS28</i> and <i>GAPDH</i> presented the most stable expression for the heat treatment. To validate the above results, a chalcone synthase gene (<i>GbCHS</i>) in ginkgo was amplified by quantitative real-time polymerase chain reaction (qRT-PCR). Our results provide different suitable reference genes for further gene expression studies in ginkgo. |
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| spelling | doaj.art-4c9aeb1311b64b0f80a9233ec937271c2023-11-20T21:32:23ZengMDPI AGForests1999-49072020-11-011111121710.3390/f11111217Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental ConditionsTingting Zhou0Xiaoming Yang1Fangfang Fu2Guibin Wang3Fuliang Cao4Co-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, ChinaCo-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, ChinaCo-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, ChinaCo-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, ChinaCo-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China<i>Ginkgo biloba</i>, a deciduous tree species in the Ginkgo family, has a long history of cultivation in China and is widely used in garden landscapes, medicine, food, and health products. However, few reports have focused on the systematic selection of optimal reference genes based on transcriptomic data in <i>G. biloba</i>. The purpose of our research was to select an internal reference gene suitable for different experimental conditions from thirteen candidate reference genes by the delta cycle threshold (ΔCt) method, geNorm, BestKeeper, NormFinder, and RefFinder programs. The reference genes were used for gene expression analyses of <i>Ginkgo biloba</i>. These results showed that elongation factor 1(<i>EF1</i>) and ubiquitin (<i>UBI)</i> were the best choices for samples of different ginkgo genotypes. The expression of <i>UBI</i> and <i>HAS28</i> presented the most stable at different developmental stages of ginkgo, and <i>EIF3I</i> and <i>RPII</i> were considered as suitable reference genes in different tissues of ginkgo. For methyl jasmonate (MeJA) treatment, <i>ACA</i> and <i>ACT</i> were identified as the optimal reference genes. For cold stress treatment, <i>RPII</i> and <i>EIF4E</i> were chosen for the gene expression normalizations. <i>HAS28</i> and <i>GAPDH</i> presented the most stable expression for the heat treatment. To validate the above results, a chalcone synthase gene (<i>GbCHS</i>) in ginkgo was amplified by quantitative real-time polymerase chain reaction (qRT-PCR). Our results provide different suitable reference genes for further gene expression studies in ginkgo.https://www.mdpi.com/1999-4907/11/11/1217<i>Ginkgo biloba</i>qRT-PCRexperimental conditionsgene expression |
| spellingShingle | Tingting Zhou Xiaoming Yang Fangfang Fu Guibin Wang Fuliang Cao Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental Conditions Forests <i>Ginkgo biloba</i> qRT-PCR experimental conditions gene expression |
| title | Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental Conditions |
| title_full | Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental Conditions |
| title_fullStr | Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental Conditions |
| title_full_unstemmed | Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental Conditions |
| title_short | Selection of Suitable Reference Genes Based on Transcriptomic Data in <i>Ginkgo biloba</i> under Different Experimental Conditions |
| title_sort | selection of suitable reference genes based on transcriptomic data in i ginkgo biloba i under different experimental conditions |
| topic | <i>Ginkgo biloba</i> qRT-PCR experimental conditions gene expression |
| url | https://www.mdpi.com/1999-4907/11/11/1217 |
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