Reference gene selection for quantitative real-time PCR normalization in Caragana intermedia under different abiotic stress conditions.
Reference gene selection for quantitative real-time PCR normalization in Caragana intermedia under different abiotic stress conditions.
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DOI：
10.1371/journal.pone.0053196
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Qi L
中科院分区：
文献类型：
--
作者：
Zhu J;Zhang L;Li W;Han S;Yang W;Qi L
定量实时逆转录聚合酶链反应（qPCR）是一种敏感的基因表达分析技术，其依赖于用于数据标准化的参考基因的稳定性。中间锦鸡儿（Caragana intermedia）是我国西部和西北部荒漠地区广泛分布的一种天然荒漠灌木，具有较强的抗旱性、固沙能力和较高的饲用价值。本研究对10个候选参考基因进行了分析。在两种不同的植物器官（根和叶）中，对不同的非生物（渗透、盐、冷和热）胁迫下的Intermedia进行了研究。使用geNorm、Normalization和BestKeeper算法评估这些基因的表达稳定性。排名最高的参考基因在不同的样品组中不同，但UNK 2，PP 2A和SAND在所有测试样品中最稳定。UNK 2和SAND将适合于标准化盐处理根的基因表达数据，而UNK 2、SAND和EF-1α的组合将适合于盐处理叶。UNK 1、UNK 2和PP 2A适合于PEG处理的（渗透）根，而TIP 41和PP 2A的组合最适合于PEG处理的叶。SAND、PP 2A和TIP 41在热处理的叶片中表现出最稳定的表达。在低温处理的叶片中，SAND和EF-1α表达最稳定。为了进一步验证本研究中鉴定的参考基因的适用性，平行研究了DREB 1和DREB 2（AtDREB 1和AtDREB 2的同源物）的表达水平。本研究首次系统地分析了在中国对虾中qPCR的上级参考基因的选择。研究了不同非生物胁迫条件下C. intermedia基因表达的变化规律，为进一步研究C. intermedia基因的表达提供了理论依据。中间锦鸡儿属（Caragana）和豆科锦鸡儿属（Caragana）的其他物种。
Quantitative real-time reverse transcription polymerase chain reaction (qPCR), a sensitive technique for gene expression analysis, depends on the stability of the reference genes used for data normalization. Caragana intermedia, a native desert shrub with strong drought-resistance, sand-fixing capacity and high forage value that is widespread in the desert land of west and northwest China, has not been investigated regarding the identification of reference genes suitable for the normalization of qPCR data. In this study, 10 candidate reference genes were analyzed in C. intermedia subjected to different abiotic (osmotic, salt, cold and heat) stresses, in two distinct plant organs (roots and leaves). The expression stability of these genes was assessed using geNorm, NormFinder and BestKeeper algorithms. The best-ranked reference genes differed across the different sets of samples, but UNK2, PP2A and SAND were the most stable across all tested samples. UNK2 and SAND would be appropriate for normalizing gene expression data for salt-treated roots, whereas the combination of UNK2, SAND and EF-1α would be appropriate for salt-treated leaves. UNK1, UNK2 and PP2A would be appropriate for PEG-treated (osmotic) roots, whereas the combination of TIP41 and PP2A was the most suitable for PEG-treated leaves. SAND, PP2A and TIP41 exhibited the most stable expression in heat-treated leaves. In cold-treated leaves, SAND and EF-1α were the most stably expressed. To further validate the suitability of the reference genes identified in this study, the expression levels of DREB1 and DREB2 (homologs of AtDREB1 and AtDREB2) were studied in parallel. This study is the first systematic analysis for the selection of superior reference genes for qPCR in C. intermedia under different abiotic stress conditions, and will benefit future studies on gene expression in C. intermedia and other species of the leguminous genus Caragana.
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DOI：
10.1073/pnas.0405313101
发表时间：
2004-12-14
影响因子：
11.1
作者：
Lo Bianco, C;Schneider, BL;Aebischer, P
通讯作者：
Aebischer, P
影响因子：
3.5
作者：
Ango, F;Robbe, D;Fagni, L
通讯作者：
Fagni, L
影响因子：
7.4
作者：
Gomez-Lazaro, Maria;Bonekamp, Nina A.;Schrader, Michael
通讯作者：
Schrader, Michael
影响因子：
120.7
作者：
Schapira, AHV;Olanow, CW
通讯作者：
Olanow, CW
影响因子：
9.9
作者：
Becker, Claudia;Jick, Susan S.;Meier, Christoph R.
通讯作者：
Meier, Christoph R.