With the availability of genome sequences, gene expression analysis of jute has drawn considerable attention for understanding the regulatory mechanisms of fiber development and improving fiber quality. Gene expression profiles of a target gene can provide valuable clues towards the understanding of its biological function. Reverse transcription quantitative real-time PCR (qRT-PCR) is the best method for targeted gene expression analysis due to its sensitivity and reproducibility. However, calculating relative expression requires reference genes, which must be stable across various biological conditions. For this purposes, 11 prospective genes namely, 28S RNA, ACT7, CYP, EF1A, EF2, ETIF3E, GAPDH, PP2Ac, PTB, UBC2 and UBI1 were evaluated for their potential use as reference genes in jute. The expression stabilities of eleven prospective genes were analyzed in various jute plant tissues, such as the root, stick, bark, leaf, flower, seed and fiber, as well as under abiotic (waterlogged, drought and salinity) and biotic stress (infestation with Macrophomina phaseolina) conditions with different time points. All 11 genes were variably expressed in different tissues and stress conditions. To find suitable reference genes in different sample sets, a comprehensive approach based on four statistical algorithms such as GeNorm, BestKeeper, NormFinder the ΔCt was used. The PP2Ac and EF2 genes were the most stably expressed across the different tissues. ACT7 and UBC2 were suitable reference genes under drought stress, and CYP and PP2Ac were the most appropriate after inoculation with Macrophomina phaseolina. Under salinity stress, PP2Ac and UBC2 were the best genes, and ACT7 and PP2Ac were the most suitable under waterlogged conditions. Expression stability of reference genes from jute varied in different tissues and selected experimental conditions. Our results provide a valuable resource for the accurate normalization of gene expression experiments in fiber research for important bast fiber crops.

中文翻译：

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黄麻实时定量 RT-PCR 分析参考基因的鉴定和验证。

随着基因组序列的获得，黄麻的基因表达分析对于了解纤维发育的调控机制和提高纤维质量引起了广泛的关注。目标基因的基因表达谱可以为理解其生物学功能提供有价值的线索。逆转录定量实时 PCR (qRT-PCR) 因其灵敏度和重现性而成为目标基因表达分析的最佳方法。然而，计算相对表达需要参考基因，该参考基因必须在各种生物条件下保持稳定。为此，评估了 11 个预期基因，即 28S RNA、ACT7、CYP、EF1A、EF2、ETIF3E、GAPDH、PP2Ac、PTB、UBC2 和 UBI1 作为黄麻参考基因的潜在用途。分析了 11 个预期基因在各种黄麻植物组织中的表达稳定性，如根、枝、皮、叶、花、种子和纤维，以及在非生物（淹水、干旱和盐度）和生物胁迫（虫害）下的表达稳定性。 Macrophomina Phaseolina）具有不同时间点的条件。所有 11 个基因在不同的组织和应激条件下都有不同的表达。为了在不同的样本集中找到合适的内参基因，使用了基于GeNorm、BestKeeper、NormFinder和ΔCt等四种统计算法的综合方法。PP2Ac 和 EF2 基因在不同组织中表达最稳定。ACT7和UBC2是干旱胁迫下合适的内参基因，CYP和PP2Ac是接种Macrophhomina Phaseolina后最合适的内参基因。在盐度胁迫下，PP2Ac和UBC2是最好的基因，ACT7和PP2Ac在淹水条件下最适合。黄麻参考基因的表达稳定性在不同组织和选定的实验条件下有所不同。我们的结果为重要麻类纤维研究中基因表达实验的准确标准化提供了宝贵的资源。