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10 November 2021 Quantification of endophyte Serendipita indica in Brassica napus roots by qPCR
Nastaran Abin, Nader Rokni, Ali Reza Shafeinia, M. Hossein Borhan
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Context. The fungal endophyte Serendipita indica enhances plant growth and plant resistance to biotic and abiotic stresses. Inoculum concentration greatly impacts the endophyte-plant interaction from mutualism to antagonism.

Aims and methods. We used both microscopy and qPCR to examine the effect of inoculum concentrations on the extent (%) and density of Brassica napus L. root colonisation by S. indica. B. napus seeds were inoculated with the fungus at five different inoculum concentrations (1-10% w/w basis).

Key results. Standard curves were constructed using the mean threshold cycle (Ct) and serially diluted gDNA ranging between 4.14 × 102 and 2.65 × 105 colony forming units (CFU). The result indicated a linear relationship between Ct and the log of input DNA. Variation in inoculum concentration significantly affected the root colonisation density by the fungus shown by qPCR. However, the percent root colonisation (PRC) measure was not affected and remained the same across all the treatments.

Conclusions. Our findings show that the qPCR assay developed will determine the colonisation density whereas PRC gives a measure of the incidence of infected roots. Also, we suggest that the optimum quantity of inoculum is a key factor for a successful interaction that impacts the plant-S. indica interaction.

Implications. To our knowledge, this is the first study that quantitative qPCR has been used to investigate the correlation between inoculum quantities and the corresponding density of root colonisation in S. indica.

© 2021 The Author(s) (or their employer(s)). Published by CSIRO Publishing.
Nastaran Abin, Nader Rokni, Ali Reza Shafeinia, and M. Hossein Borhan "Quantification of endophyte Serendipita indica in Brassica napus roots by qPCR," Crop and Pasture Science 72(12), 985-993, (10 November 2021).
Received: 30 December 2020; Accepted: 16 August 2021; Published: 10 November 2021
absolute quantification
inoculation concentration
inoculum density
molecular detection
plant growth promotion
plant-endophyte interaction
real-time PCR
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