Selection of reference genes for expression profiling in biostimulation research of soybean

IF 5.2 2区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Chemical and Biological Technologies in Agriculture Pub Date : 2024-09-03 DOI:10.1186/s40538-024-00660-3

Magdalena Sozoniuk, Michał Świeca, Andrea Bohatá, Petr Bartoš, Jan Bedrníček, František Lorenc, Markéta Jarošová, Kristýna Perná, Adéla Stupková, Jana Lencová, Pavel Olšan, Jan Bárta, Agnieszka Szparaga, María Cecilia Pérez-Pizá, Sławomir Kocira

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Abstract

Background

Plant biostimulants constitute a promising environmentally friendly alternative for increasing crop yield and tolerance to unfavorable conditions. Among various types of such formulations, botanical extracts are gaining more recognition as products supporting plant performance. Moreover, novel tools such as cold-plasma or low-pressure microwave plasma discharge are being proposed as techniques that might improve their efficacy. Elucidation of the biostimulant’s mode of action requires complex research at a molecular level. Transcriptional changes occurring after biostimulant spraying might be investigated using RT-qPCR. However, this technique requires data normalization against stable endogenous controls.

Results

Here, we tested the expression stability of ten candidate genes in soybean plants exposed to various biostimulants treatment. Selection of the best-performing reference genes was conducted using four algorithms (geNorm, NormFinder, BestKeeper, and ΔCt method). According to the obtained results, Bic-C2 (RNA-binding protein Bicaudal-C) and CYP (cyclophilin type peptidyl-prolyl cis–trans isomerase) showed highest expression stability, while expression of EF1B (elongation factor 1-beta) fluctuated the most among a tested set of candidate genes.

Conclusions

Overall, we recommend using Bic-C2 together with CYP for the RT-qPCR data normalization in soybean biostimulation experiments. To our best knowledge, this is the first comprehensive study of reference genes stability in plants subjected to biostimulant treatment. The results of this study will aid in further biostimulant research in crop plants, facilitating analyses performed on the transcriptional level.

Graphical Abstract

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大豆生物刺激研究中表达谱分析参考基因的选择

背景植物生物刺激剂是一种很有前途的环境友好型替代品，可提高作物产量和对不利条件的耐受性。在各种类型的此类制剂中，植物提取物作为支持植物生长的产品正得到越来越多的认可。此外，冷等离子体或低压微波等离子体放电等新型工具也被认为是可以提高其功效的技术。阐明生物刺激剂的作用模式需要在分子水平上进行复杂的研究。喷洒生物刺激剂后发生的转录变化可通过 RT-qPCR 进行研究。结果在这里，我们检测了 10 个候选基因在大豆植株暴露于各种生物刺激剂处理后的表达稳定性。我们使用四种算法（geNorm、NormFinder、BestKeeper 和 ΔCt 法）选出了表现最佳的参考基因。结果表明，Bic-C2（RNA 结合蛋白 Bicaudal-C）和 CYP（环嗜素型肽基-脯氨酰顺反异构酶）的表达稳定性最高，而 EF1B（伸长因子 1-beta）的表达在一组候选基因中波动最大。据我们所知，这是首次对接受生物刺激剂处理的植物中参考基因的稳定性进行全面研究。这项研究的结果将有助于在作物植物中进一步开展生物刺激剂研究，促进转录水平的分析。

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来源期刊

Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology

CiteScore

6.80

自引率

3.00%

发文量

审稿时长

15 weeks

期刊介绍： Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.