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
{"title":"大豆生物刺激研究中表达谱分析参考基因的选择","authors":"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","doi":"10.1186/s40538-024-00660-3","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><p>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.</p><h3>Results</h3><p>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, <i>Bic-C2</i> (RNA-binding protein Bicaudal-C) and <i>CYP</i> (cyclophilin type peptidyl-prolyl cis–trans isomerase) showed highest expression stability, while expression of <i>EF1B</i> (elongation factor 1-beta) fluctuated the most among a tested set of candidate genes.</p><h3>Conclusions</h3><p>Overall, we recommend using <i>Bic-C2</i> together with <i>CYP</i> 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. 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Selection of reference genes for expression profiling in biostimulation research of soybean
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.
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.
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