Chauhan Abhishek Singh, Tiwari Madhu, Indoliya Yuvraj, Mishra Shashank Kumar, Lavania Umesh Chandra, Chauhan Puneet Singh, Chakrabarty Debasis, Tripathi Rudra Deo
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002 India.
Molecular Biology and Biotechnology Division, CSIR - National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001 India.
Physiol Mol Biol Plants. 2023 May;29(5):613-627. doi: 10.1007/s12298-023-01315-7. Epub 2023 May 25.
Vetiver [ (L.) Roberty] is a perennial C-4 grass traditionally valued for its aromatic roots/root essential oil. Owing to its deep penetrating web-forming roots, the grass is now widely used across the globe for phytoremediation and the conservation of soil and water. This study has used the transcriptome data of vetiver roots in its two distinct geographic morphotypes (North Indian type A and South Indian type B) for reference gene(s) identification. Further, validation of reference genes using various abiotic stresses such as heat, cold, salt, and drought was carried out. The de novo assembly based on differential genes analysis gave 1,36,824 genes (PRJNA292937). Statistical tests like RefFinder, NormFinder, BestKeeper, geNorm, and Delta-Ct software were applied on 346 selected contigs. Eleven selected genes viz., , , , , along with were used for qRT-PCR analysis. Finally, the study identified the five best reference genes and along with . The two optimal reference genes and were identified with the help of qbase + software. The findings of the present analyses have value in the identification of suitable reference gene(s) in transcriptomic and molecular data analysis concerning various phenotypes related to abiotic stress and developmental aspects, as well as a quality control measure in gene expression experiments. Identifying reference genes in vetiver appears important as it allows for accurate normalization of gene expression data in qRT-PCR experiments.
The online version contains supplementary material available at 10.1007/s12298-023-01315-7.
岩兰草[(L.)罗伯蒂]是一种多年生C4草本植物,其芳香根/根精油传统上备受珍视。由于其根系深入且形成网状,这种草现在在全球广泛用于植物修复以及水土保持。本研究利用了岩兰草两种不同地理形态类型(北印度A型和南印度B型)根系的转录组数据来鉴定参考基因。此外,还利用热、冷、盐和干旱等各种非生物胁迫对参考基因进行了验证。基于差异基因分析的从头组装得到了136824个基因(PRJNA292937)。对346个选定的重叠群应用了RefFinder、NormFinder、BestKeeper、geNorm和Delta-Ct软件等统计测试。11个选定的基因,即 、 、 、 、 以及 用于qRT-PCR分析。最后,该研究鉴定出了五个最佳参考基因 、 以及 。借助qbase +软件鉴定出了两个最佳参考基因 和 。本分析的结果对于在与非生物胁迫和发育方面相关的各种表型的转录组学和分子数据分析中鉴定合适的参考基因具有价值,同时也是基因表达实验中的一种质量控制措施。在岩兰草中鉴定参考基因似乎很重要,因为它能在qRT-PCR实验中实现基因表达数据的准确标准化。
在线版本包含可在10.1007/s12298-023-01315-7获取的补充材料。