Practice Abstract - Research and innovation

Absolute qPCR quantification of arbuscular mycorrhiza fungi in crop plants

Problem

Conventional methods to quantify arbuscular mycorrhizal fungal (AMF) colonization include microscopy, which involves staining of the roots and meticulous observation. This method is time consuming and subjective because it depends on the observer’s level of expertise.

Solution

The use of broad range qPCR primers was validated to quantify AMF root colonization in different crops. This high-throughput and reliable technique is versatile and suitable for a wide range of research projects.

Benefits and weaknesses

Compared to the conventional microscopy method, qPCR allows the simultaneous analysis of a larger number of samples. Moreover, the results of qPCR are not influenced by observer subjectivity. AMF quantification can be complemented by sequencing the PCR product to assess AMF diversity. One weakness of the qPCR method is that it cannot distinguish between the different AMF  structures. In addition, it is worth noting that the qPCR method
yielded inconclusive results when applied to leek, possibility due to negligible changes in colonization rate (Corona Ramírez et al. 2023). We conclude that the qPCR method should be validated for each crop.

For more information, please see https://zenodo.org/records/10812217

Source Project
Root phenotyping and genetic improvement for rotational crops resilient to environmental change
Ongoing | 2022-2027
Main funding source
Horizon Europe (EU Research and Innovation Programme)
Geographical location
France, Slovenia, Spain, Italy, Ireland, Germany, Netherlands, Denmark, Austria, United Kingdom, Switzerland, Morocco, South Africa
Project details