Relative qPCR to quantify colonization of plant roots by arbuscular mycorrhizal fungi
| Autor: | Natacha Bodenhausen, Marcel G. A. van der Heijden, Alain Y. Held, Klaus Schlaeppi, Gabriel Deslandes-Hérold, Jan Waelchli |
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| Přispěvatelé: | University of Zurich, Schlaeppi, Klaus |
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Evolution Soil biology Arbuscular mycorrhizal fungi Plant Science 580 Plants (Botany) Relative quantification Plant Roots 01 natural sciences Petunia Soil 03 medical and health sciences 10126 Department of Plant and Microbial Biology 1311 Genetics Behavior and Systematics Mycorrhizae 1110 Plant Science Botany 1312 Molecular Biology Genetics Colonization 10211 Zurich-Basel Plant Science Center Molecular Biology Soil Microbiology Ecology Evolution Behavior and Systematics 030304 developmental biology Quantitative PCR (qPCR) 0303 health sciences Ecology biology Plant roots Host (biology) fungi Fungi General Medicine Plants Amplicon biology.organism_classification Arbuscular mycorrhiza 1105 Ecology Evolution Behavior and Systematics Crop combinations and interactions Original Article 010606 plant biology & botany |
| Zdroj: | Mycorrhiza Bodenhausen, Natacha; Deslandes-Hérold, Gabriel; Waelchli, Jan; Held, Alain; van der Heijden, Marcel G. A.; Schlaeppi, Klaus (2021). Relative qPCR to quantify colonization of plant roots by arbuscular mycorrhizal fungi. Mycorrhiza, 31(2), pp. 137-148. Springer 10.1007/s00572-020-01014-1 |
| ISSN: | 1432-1890 0940-6360 |
| DOI: | 10.1007/s00572-020-01014-1 |
| Popis: | Arbuscular mycorrhiza fungi (AMF) are beneficial soil fungi that can promote the growth of their host plants. Accurate quantification of AMF in plant roots is important because the level of colonization is often indicative of the activity of these fungi. Root colonization is traditionally measured with microscopy methods which visualize fungal structures inside roots. Microscopy methods are labor-intensive, and results depend on the observer. In this study, we present a relative qPCR method to quantify AMF in which we normalized the AMF qPCR signal relative to a plant gene. First, we validated the primer pair AMG1F and AM1 in silico, and we show that these primers cover most AMF species present in plant roots without amplifying host DNA. Next, we compared the relative qPCR method with traditional microscopy based on a greenhouse experiment with Petunia plants that ranged from very high to very low levels of AMF root colonization. Finally, by sequencing the qPCR amplicons with MiSeq, we experimentally confirmed that the primer pair excludes plant DNA while amplifying mostly AMF. Most importantly, our relative qPCR approach was capable of discriminating quantitative differences in AMF root colonization and it strongly correlated (Spearman Rho = 0.875) with quantifications by traditional microscopy. Finally, we provide a balanced discussion about the strengths and weaknesses of microscopy and qPCR methods. In conclusion, the tested approach of relative qPCR presents a reliable alternative method to quantify AMF root colonization that is less operator-dependent than traditional microscopy and offers scalability to high-throughput analyses. |
| Databáze: | OpenAIRE |
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