Lucho Simone Ribeiro, do Amaral Marcelo Nogueira, Benitez Letícia Carvalho, Milech Cristini, Kleinowski Alitcia Moraes, Bianchi Valmor João, Braga Eugenia Jacira Bolacel
Department of Botany, Federal University of Pelotas, Pelotas, RS Brazil.
Physiol Mol Biol Plants. 2018 Sep;24(5):767-779. doi: 10.1007/s12298-018-0583-7. Epub 2018 Jul 25.
is an important source of natural steviol glycosides and is of increasing interest in various fields of study. Therefore, understanding the molecular processes regulating its metabolism is of great importance. In this study, the stability of seven reference genes (- and ) under the effect of three stress-related elicitors (methyl jasmonate, salicylic acid, and spermidine) was evaluated in stevia plants. We used RefFinder software, which makes use of the four main currently available algorithms for reference gene selection: geNorm, NormFinder, BestKeeper, and the Comparative ∆Ct method. The results indicated that and can be used as reference genes under all tested experimental conditions. The genes, traditionally used as a reference gene, along with , showed the lowest stability. The expression of -- and genes was used to confirm the validated reference genes, showing that inadequacy of the reference gene may lead to erroneous results. This is the first study on the stability of reference genes in plants, and is of great relevance for further analysis of the gene expression of the steviol glycoside biosynthetic pathway.
是天然甜菊糖苷的重要来源，并且在各个研究领域中越来越受到关注。因此，了解调节其代谢的分子过程非常重要。在本研究中，评估了七种参考基因（- 和 ）在三种与胁迫相关的诱导剂（茉莉酸甲酯、水杨酸和亚精胺）作用下在甜叶菊植物中的稳定性。我们使用了RefFinder软件，该软件利用目前可用的四种主要参考基因选择算法：geNorm、NormFinder、BestKeeper和比较∆Ct法。结果表明， 和 在所有测试实验条件下都可作为参考基因。传统上用作参考基因的基因，以及 ，稳定性最低。使用-- 和 基因的表达来确认经过验证的参考基因，表明参考基因的不足可能导致错误的结果。这是关于甜叶菊植物中参考基因稳定性的首次研究，对于进一步分析甜菊糖苷生物合成途径的基因表达具有重要意义。
