Validation of potential reference genes for qPCR in maize across abiotic stresses, hormone treatments, and tissue types.
Validation of potential reference genes for qPCR in maize across abiotic stresses, hormone treatments, and tissue types.
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在非生物胁迫、激素处理和组织类型中验证玉米 qPCR 的潜在参考基因
DOI：
10.1371/journal.pone.0095445
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Zhang S
中科院分区：
文献类型：
--
作者：
Lin Y;Zhang C;Lan H;Gao S;Liu H;Liu J;Cao M;Pan G;Rong T;Zhang S
逆转录定量聚合酶链反应（RT-qPCR）是一种功能强大且应用广泛的基因表达检测技术。内参基因作为内源性控制，确保结果的准确性和可重复性，对数据规范化至关重要。为了支持玉米内参基因选择的文献，我们评估了10个候选内参基因，包括8个传统的内参基因和2个来自微阵列数据集的潜在候选基因，在非生物胁迫（冷、热、盐和PEG）、植物激素处理（脱落酸、水杨酸、茉莉酸、乙烯和赤霉素）和不同组织类型下玉米的表达水平。使用geNorm、NormFinder和Bestkeeper三个分析软件包评估内参基因表达的稳定性。结果表明，延长因子1α (EF1α)、微管蛋白β (β-TUB)、亲环蛋白（CYP）和真核起始因子4A （EIF4A）是玉米基因整体表达规范化最可靠的内参基因，而GRP（富甘氨酸rna结合蛋白）、GLU1（β-葡萄糖苷酶）和UBQ9（泛素9）是最不稳定和最不适合的内参基因。此外，通过验证WRKY50在不同样品中的表达，确认EF1α、β-TUB及其组合作为内参基因的适宜性。目前的研究为玉米在某些非生物胁迫、激素和组织类型中基因表达正常化的迫切需求提供了合适的内参基因。
The reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a powerful and widely used technique for the measurement of gene expression. Reference genes, which serve as endogenous controls ensure that the results are accurate and reproducible, are vital for data normalization. To bolster the literature on reference gene selection in maize, ten candidate reference genes, including eight traditionally used internal control genes and two potential candidate genes from our microarray datasets, were evaluated for expression level in maize across abiotic stresses (cold, heat, salinity, and PEG), phytohormone treatments (abscisic acid, salicylic acid, jasmonic acid, ethylene, and gibberellins), and different tissue types. Three analytical software packages, geNorm, NormFinder, and Bestkeeper, were used to assess the stability of reference gene expression. The results revealed that elongation factor 1 alpha (EF1α), tubulin beta (β-TUB), cyclophilin (CYP), and eukaryotic initiation factor 4A (EIF4A) were the most reliable reference genes for overall gene expression normalization in maize, while GRP (Glycine-rich RNA-binding protein), GLU1(beta-glucosidase), and UBQ9 (ubiquitin 9) were the least stable and most unsuitable genes. In addition, the suitability of EF1α, β-TUB, and their combination as reference genes was confirmed by validating the expression of WRKY50 in various samples. The current study indicates the appropriate reference genes for the urgent requirement of gene expression normalization in maize across certain abiotic stresses, hormones, and tissue types.
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影响因子：
5.3
作者：
Artico S;Nardeli SM;Brilhante O;Grossi-de-Sa MF;Alves-Ferreira M
通讯作者：
Alves-Ferreira M
影响因子：
6.9
作者：
Hartung, W;Sauter, A;Hose, E
通讯作者：
Hose, E
影响因子：
--
作者：
Hu R;Fan C;Li H;Zhang Q;Fu YF
通讯作者：
Fu YF
影响因子：
3.7
作者：
Chandna R;Augustine R;Bisht NC
通讯作者：
Bisht NC
影响因子：
5.2
作者：
Lin, Y. L.;Lai, Z. X.
通讯作者：
Lai, Z. X.