Quantification of endophyte Serendipita indica in Brassica napus roots by qPCR
Nastaran Abin A D , Nader Rokni
B * , Ali Reza Shafeinia A D and M. Hossein Borhan C
+ Author Affiliations
- Author Affiliations
A Faculty of Agriculture, Department of Biotechnology, Ramin Agricultural and Natural Resources University, PO Box 73637-63417, Ahwaz, Iran.
B Faculty of Science, Jundi-Shapur University of Technology, PO Box 64615-334, Dezful, Iran.
C Saskatoon Research Centre, Agriculture and Agri-Food Canada, Saskatoon, SK, Canada.
D Department of Plant Production Engineering and Genetics, Agricultural Sciences and Natural Resources University of Khusetan, PO Box 73637-63417, Ahwaz, Iran.
Crop & Pasture Science 72(12) 985-993 https://doi.org/10.1071/CP21265
Submitted: 30 December 2020 Accepted: 16 August 2021 Published: 10 November 2021
© 2021 The Author(s) (or their employer(s)). Published by CSIRO Publishing
Abstract
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.
Keywords: absolute quantification, inoculation concentration, inoculum density, molecular detection, plant-endophyte interaction, plant growth promotion, real-time PCR, symbiosis.
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