| 红掌佛焰苞基因qRT-PCR分析中内参基因的筛选 |
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| 引用本文： | 杨澜,杨光穗,李崇晖,牛俊海,尹俊梅.红掌佛焰苞基因qRT-PCR分析中内参基因的筛选[J].热带亚热带植物学报,2015,23(1):51-58. |
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| 作者姓名： | 杨澜 杨光穗 李崇晖 牛俊海 尹俊梅 |
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| 作者单位： | 1. 中国热带农业科学院热带作物品种资源研究所，农业部华南作物基因资源与种质创制重点实验室，海南 儋州 571737; 贵州省园艺研究所，贵阳 550006
2. 中国热带农业科学院热带作物品种资源研究所，农业部华南作物基因资源与种质创制重点实验室，海南 儋州 571737 |
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| 基金项目： | 国家自然科学基金项目(31101578, 31201652); 海南省重大科技项目(ZDZX2013012);中央级公益性科研院所基本科研业务费专项(1630032012017)资助 |
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| 摘 要： | 为筛选红掌(Anthurium andraeanumLinden)中稳定表达、可用于佛焰苞中实时荧光定量PCR分析(qRT-PCR)的内参基因，对5个组成型表达基因EF1-a、UBQ7、ACTB、GADPH、His3进行表达稳定性分析，并利用所筛选的内参基因研究红掌的二氢黄酮醇还原酶基因(dfr)的表达水平。结果表明，5种内参基因在不同品种间的表达稳定性不同。据内参基因标准化因子的配对差异分析(Vn/n+1)，判定内参基因的最适数目为2，ACTB和UBQ7在红掌不同品种及佛焰苞发育不同阶段中表达均稳定，是理想的内参基因。dfr在不同品种的佛焰苞及佛焰苞发育过程中均有表达，且dfr表达水平的变化趋势一致，因此，所选内参基因是合适的。
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| 关 键 词： | 红掌 佛焰苞 实时荧光定量PCR 内参基因 |
| 收稿时间： | 2014/4/16 0:00:00 |
| 修稿时间： | 2014/9/11 0:00:00 |
Screening of Reference Genes in Anthurium andraeanum Spathes for qRT-PCR Analysis |
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| YANG Lan,YANG Guang-sui,LI Chong-hui,NIU Jun-hai and YIN Jun-mei.Screening of Reference Genes in Anthurium andraeanum Spathes for qRT-PCR Analysis[J].Journal of Tropical and Subtropical Botany,2015,23(1):51-58. |
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| Authors: | YANG Lan YANG Guang-sui LI Chong-hui NIU Jun-hai and YIN Jun-mei |
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| Institution: | 1. Institute of Tropical Crops Genetic Resources, Chinese Academy of Tropical Agricultural Sciences, Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture, Danzhou 571737, China;2. Horticultural Research Institute of Guizhou Province, Guiyang 550006, China,Institute of Tropical Crops Genetic Resources, Chinese Academy of Tropical Agricultural Sciences, Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture, Danzhou 571737, China,Institute of Tropical Crops Genetic Resources, Chinese Academy of Tropical Agricultural Sciences, Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture, Danzhou 571737, China,Institute of Tropical Crops Genetic Resources, Chinese Academy of Tropical Agricultural Sciences, Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture, Danzhou 571737, China and Institute of Tropical Crops Genetic Resources, Chinese Academy of Tropical Agricultural Sciences, Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture, Danzhou 571737, China |
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| Abstract: | In order to screening stable reference genes that can be used to real-time quantitative PCR (qRT-PCR) analysis in Anthurium andraeanum spathes, five constitutively expressed genes, including EF1-a, UBQ7, ACTB, GADPH and His3, were analyzed expression stability by using qRT-PCR, and the expression of dihydroflavonol 4-reductase gene (dfr) was analyzed by using screened reference gene. The results showed that the expression stabilities of five reference genes were different among different cultivars. On the basis of the normalization factor Vn/n+1, the optimum number of reference gene was two. The expression of ACTB and UBQ7 were the most stable in dfr ferent cultivars and different development stages of A. andraeanum spathes, so that they were ideal reference genes. When ACTB and UBQ7 were used as reference genes, the gene dfr expressed in different cultivars and different development stages of A. andraeanum spathes, and the change trend of dfr expression were consistent. Thus, both ACTB and UBQ7 were suitable as reference genes for A. andraeanum spathes. |
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| Keywords: | Anthunium andraenum Spathe qRT-PCR Reference gene |
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