Yu Mingjun, Jin Jiarui, Li Xiaoli, Liu Yuping, Su Xu, Caraballo-Ortiz Marcos A, Zhang Penghui, Yang Qian, Qu Rongju, Cairang Zhaxi, Gao Xuanlin, Zheng Yinghui
School of Life Sciences, Qinghai Normal University, Xining, 810008, China.
School of Life Sciences, Qinghai Normal University, Xining, 810008, China; Key Laboratory of Biodiversity Formation Mechanism and Comprehensive Utilization of the Qinghai-Tibet Plateau in Qinghai Province, Qinghai Normal University, Xining, 810008, China; Academy of Plateau Science and Sustainability, Qinghai Normal University, Xining, 810016, China.
J Plant Physiol. 2025 Feb;305:154435. doi: 10.1016/j.jplph.2025.154435. Epub 2025 Jan 17.
Rheum tanguticum, an endemic species from the Qinghai-Xizang Plateau, is a significant perennial and medicinal plant recognized for its robust resistance to abiotic stresses, including drought, cold, and salinity. To advance the understanding of stress-response mechanisms in R. tanguticum, this study aimed to establish a reliable set of housekeeping genes as references for normalizing RT-qPCR gene expression analyses. Ten candidate genes were selected based on prior studies of related species and transcriptomic data for R. tanguticum. Their expression stability was evaluated across three tissue types (leaf, stem, and root) under four abiotic stress treatments using a comprehensive suite of bioinformatics tools. Our findings revealed that RtEF1α was the most stable reference gene in salt- and drought-stressed roots and drought-stressed stems, while RtTUB was most stable in salt-stressed stems. RtSAND was identified as the most stable reference gene in salt-, drought-, heavy metal-, and hormone-stressed leaves. For heavy metal- and hormone-stressed roots and heavy metal-stressed stems, RtUBC demonstrated the highest stability, whereas RteIF6A was most stable in hormone-stressed stems. Furthermore, the utility of these reference genes was validated by assessing the expression pattern of the drought-related gene RtNAC. This study is the first report on selecting and validating reference genes of R. tanguticum under various stress conditions, which will benefit future investigations of the genomic mechanisms involved on stress resistance in this species.
唐古特大黄是一种来自青藏高原的特有物种，是一种重要的多年生药用植物，以其对包括干旱、寒冷和盐度在内的非生物胁迫具有强大的抗性而闻名。为了深入了解唐古特大黄的胁迫响应机制，本研究旨在建立一组可靠的管家基因作为标准化RT-qPCR基因表达分析的参考。基于对相关物种的先前研究和唐古特大黄的转录组数据，选择了10个候选基因。使用一套综合的生物信息学工具，在四种非生物胁迫处理下，对三种组织类型（叶、茎和根）中的它们的表达稳定性进行了评估。我们的研究结果表明，RtEF1α是盐胁迫和干旱胁迫下的根以及干旱胁迫下的茎中最稳定的参考基因，而RtTUB在盐胁迫下的茎中最稳定。RtSAND被确定为在盐胁迫、干旱胁迫、重金属胁迫和激素胁迫下的叶中最稳定的参考基因。对于重金属胁迫和激素胁迫下的根以及重金属胁迫下的茎，RtUBC表现出最高的稳定性，而RteIF6A在激素胁迫下的茎中最稳定。此外，通过评估干旱相关基因RtNAC的表达模式，验证了这些参考基因的实用性。本研究是关于在各种胁迫条件下选择和验证唐古特大黄参考基因的首次报道，这将有利于未来对该物种抗逆基因组机制的研究。
