Cowpea and abiotic stresses: identification of reference genes for transcriptional profiling by qPCR.
Cowpea and abiotic stresses: identification of reference genes for transcriptional profiling by qPCR.
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DOI：
10.1186/s13007-018-0354-z
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发表时间：
2018
期刊：
影响因子：
5.1
通讯作者：
Benko-Iseppon AM
中科院分区：
文献类型：
--
作者：
Amorim LLB;Ferreira-Neto JRC;Bezerra-Neto JP;Pandolfi V;de Araújo FT;da Silva Matos MK;Santos MG;Kido EA;Benko-Iseppon AM
由于豇豆在贫瘠土壤中固氮的能力以及对干旱和盐胁迫的相对耐受性，人们一直致力于鉴定赋予该物种胁迫耐受性的基因和途径。实时定量PCR（qPCR）因其高准确性和特异性，已被广泛应用于基因表达的检测。在本研究中，9个候选参考基因进行了严格的测试，他们的应用程序在规范化的qPCR数据到根的四个不同的豇豆加入下两个非生物胁迫：根脱水和盐（NaCl，100 mM）。此外，在相同的条件下，四个目标转录本的调节也被仔细检查。 geNorm、Normandy、BestKeeper和ΔCt方法的结果表明，每种胁迫条件下都有一组三个经统计学验证的RG：（I）根脱水（肌动蛋白、泛素缀合酶E2变体1D和菜豆未知基因UNK），（II）盐（泛素缀合酶E2变体1D、F-box蛋白和UNK）。目标转录本的表达谱表明，类黄酮是重要的球员在豇豆响应非生物胁迫分析，因为查尔酮异构酶和查尔酮合成酶上调的宽容和敏感的加入。脂质转运蛋白也参与了豇豆对根脱水和盐胁迫的耐受机制。所述转录本在两个耐受性材料中上调，并且在敏感性材料中没有差异表达。几丁质酶B是植物在盐胁迫下的重要靶基因，在耐盐材料中表达上调，而在敏感材料中表达下调。鉴定了在根脱水和盐胁迫下豇豆中适用于qPCR分析的参考基因。这一行动将导致对该物种的基因表达进行更准确和可靠的分析。此外，在这项研究中获得的结果可能会指导未来的研究基因表达在豇豆下的其他非生物胁迫类型，施加渗透不平衡。分析的靶基因，反过来，值得功能评估，由于其转录调控压力和生物技术潜力。本文的在线版本（10.1186/s13007-018-0354-z）包含补充材料，可供授权用户使用。
Due to cowpea ability to fix nitrogen in poor soils and relative tolerance to drought and salt stresses, efforts have been directed to identifying genes and pathways that confer stress tolerance in this species. Real-time quantitative PCR (qPCR) has been widely used as the most reliable method to measure gene expression, due to its high accuracy and specificity. In the present study, nine candidate reference genes were rigorously tested for their application in normalization of qPCR data onto roots of four distinct cowpea accessions under two abiotic stresses: root dehydration and salt (NaCl, 100 mM). In addition, the regulation of four target transcripts, under the same referred conditions was also scrutinized. geNorm, NormFinder, BestKeeper, and ΔCt method results indicated a set of three statistically validated RGs for each stress condition: (I) root dehydration (actin, ubiquitin-conjugating enzyme E2 variant 1D, and a Phaseolus vulgaris unknown gene—UNK), and (II) salt (ubiquitin-conjugating enzyme E2 variant 1D, F-box protein, and UNK). The expression profile of the target transcripts suggests that flavonoids are important players in the cowpea response to the abiotic stresses analyzed, since chalcone isomerase and chalcone synthase were up-regulated in the tolerant and sensitive accessions. A lipid transfer protein also participates in the cowpea tolerance mechanisms to root dehydration and salt stress. The referred transcript was up-regulated in the two tolerant accessions and presented no differential expression in the sensitive counterparts. Chitinase B, in turn, generally related to plant defense, was an important target transcript under salt stress, being up-regulated at the tolerant, and down-regulated in the sensitive accession. Reference genes suitable for qPCR analyses in cowpea under root dehydration and salt stress were identified. This action will lead to a more accurate and reliable analysis of gene expression on this species. Additionally, the results obtained in this study may guide future research on gene expression in cowpea under other abiotic stress types that impose osmotic imbalance. The target genes analyzed, in turn, deserve functional evaluation due to their transcriptional regulation under stresses and biotechnological potential. The online version of this article (10.1186/s13007-018-0354-z) contains supplementary material, which is available to authorized users.
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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：
5.1
作者：
Caldana C;Scheible WR;Mueller-Roeber B;Ruzicic S
通讯作者：
Ruzicic S
DOI：
10.1007/978-1-4614-0634-1_9
发表时间：
2012-01-01
期刊：
ABIOTIC STRESS RESPONSES IN PLANTS: METABOLISM, PRODUCTIVITY AND SUSTAINABILITY
影响因子：
--
作者：
Di Ferdinando, Martina;Brunetti, Cecilia;Tattini, Massimiliano
通讯作者：
Tattini, Massimiliano
影响因子：
5.3
作者：
Exposito-Rodriguez, Marino;Borges, Andres A.;Borges-Perez, Andres;Perez, Jose A.
通讯作者：
Perez, Jose A.
影响因子：
3.7
作者：
Zhu J;Zhang L;Li W;Han S;Yang W;Qi L
通讯作者：
Qi L