Experimental validation under controlled conditions of real time PCR to quantify arbuscular mycorrhizal colonization in root

Arruda, Bruna; Rodrigues, Yasmin Florentino; Herrera, Wilfrand Ferney Bejarano; Robin, Agnès; Cotta, Simone Raposo; Andreote, Fernando Dini

doi:10.1016/j.mimet.2021.106382

Cited by 5 publications

(2 citation statements)

References 16 publications

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“…It is worth noting that in this study we estimated AM fungal colonisation by quantifying the ratio of DNA copy numbers between arbuscular mycorrhizal fungi and roots. This molecular quantification method of AM fungal colonization has been used in an increasing number of studies (Arruda et al., 2022; Bodenhausen et al., 2021; Duret et al., 2022) as it may require less expertise than conventional microscopy. As a support to this methodology, we found a significant (but not very strong) correlation between molecular and microscopic quantification of AM fungal colonisation among a subset of our species ( r = 0.46, p = 0.04, Figure S4).…”

Section: Discussionmentioning

confidence: 99%

“…Across AM plant species, the abundance of AM fungi in absorptive roots was estimated by quantifying the DNA copy number of AM fungi relative to the DNA copy number of root tissue (Arruda et al, 2022;Bodenhausen et al, 2021;Duret et al, 2022). We used primers AMG1F (ATAGG GAT AGT TGG GGGCAT) and AM1 (GTTTC CCG TAA GGC GCCGAA) (Hewins et al, 2015) to quantify AM fungi, and primers trnL-g (GGGCA ATC CTG AGC CAA) and trnL-h (CCATT GAG TCT CTG CAC CTATC) to quantify the roots (Taberlet et al, 2007).…”

Section: Root Trait Measurementsmentioning

confidence: 99%

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Positioning absorptive root respiration in the root economics space across woody and herbaceous species

Liang,

Guo,

McCormack

et al. 2023

Journal of Ecology

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Abstract1. Root respiration is essential for nutrient acquisition. The respiration rate of absorptive roots theoretically relates to the economics of carbon‐nutrient exchange, but its empirical role remains largely unexplored in the trait space defining nutrient uptake strategies.2. Here, we measured the respiration rates of the distal, non‐woody, absorptive roots of 252 woody and herbaceous species from subtropical and temperate climate zones, including both arbuscular mycorrhizal and ectomycorrhizal fungal hosts.3. We found a consistent and positive correlation between root respiration rate and specific root length (root length per dry weight), irrespective of growth form, mycorrhizal type and climate zone. Root respiration rate was also positively, but less strongly and less frequently correlated with root nitrogen concentration. Root morphology strongly explained the fast‐slow gradient of root respiration in the root economics space.4. By quantifying the ratio of arbuscular mycorrhizal fungal DNA copy number and root tissue DNA copy number using qPCR, we found that the morphology‐driven gradient did not explain the full variation in fungal collaboration; thick roots were consistently well colonised, but medium and thin roots displayed a wide range of colonisation intensity.5. Synthesis: These results advance our understanding of the fundamental trait relationships that underpin the root economics space. Our study also provides a physiological linkage to the frequently measured root morphological traits and relates the root economics space to root‐derived carbon‐nutrient cycling processes.

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Section: Discussionmentioning

confidence: 99%

Section: Root Trait Measurementsmentioning

confidence: 99%

Positioning absorptive root respiration in the root economics space across woody and herbaceous species

Liang,

Guo,

McCormack

et al. 2023

Journal of Ecology

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Quantification of arbuscular mycorrhizal fungi root colonization in wheat, tomato, and leek using absolute qPCR

Corona Ramírez,

Symanczik,

Gallusser

et al. 2023

Mycorrhiza

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Arbuscular mycorrhizal fungi (AMF) form symbioses with most terrestrial plants and are known to have a positive effect on plant growth and health. Different methodologies have been developed to assess the AMF-plant symbiosis. The most applied method, which involves staining of roots and microscopic observation of the AMF structures, is tedious and time-consuming and the results are highly dependent on the observer. Using quantitative polymerase chain reaction (qPCR) to quantify AMF root colonization represents a reliable, high-throughput technique that allows the assessment of numerous samples. Quantification with qPCR can be performed through two methods: relative quantification and absolute quantification. In relative quantification, the target gene is normalized with a reference gene. On the other hand, absolute quantification involves the use of a standard curve, for which template DNA is serially diluted. In a previous paper, we validated the primer pair AMG1F and AM1 for a relative quantification approach to assess AMF root colonization in Petunia. Here, we tested the same primers with an absolute quantification approach and compared the results with the traditional microscopy method. We evaluated the qPCR method with three different crops, namely, wheat (cv. Colmetta and Wiwa), tomato, and leek. We observed a strong correlation between microscopy and qPCR for Colmetta (r = 0.90, p < 0.001), Wiwa (r = 0.94, p < 0.001), and tomato (r = 0.93, p < 0.001), but no correlation for leek (r = 0.27, p = 0.268). This highlights the importance of testing the primer pair for each specific crop.

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Can cardiolipins be used as a biomarker for arbuscular mycorrhizal fungi?

Řezanka,

Hršelová,

Kyselová

et al. 2023

Mycorrhiza

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Experimental validation under controlled conditions of real time PCR to quantify arbuscular mycorrhizal colonization in root

Cited by 5 publications

References 16 publications

Positioning absorptive root respiration in the root economics space across woody and herbaceous species

Positioning absorptive root respiration in the root economics space across woody and herbaceous species

Quantification of arbuscular mycorrhizal fungi root colonization in wheat, tomato, and leek using absolute qPCR

Can cardiolipins be used as a biomarker for arbuscular mycorrhizal fungi?

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