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Field application of a DNA-based assay to the measurement of roots of perennial grasses

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Abstract

Background and aims

DNA-based methods present new opportunities for overcoming the difficulties of accurately identifying and quantifying roots of different plant species in field soils. In order to quantify species-specific root biomass from measurements of DNA, consideration needs to be given to replication and ability to recover roots for calibration purposes in order to account for spatial, temporal and inter- and intra-species variation in DNA content of roots and distribution of roots within the soil profile.

Methods

This paper develops the field application of a DNA-based technique for direct quantification of roots in soils. The method was applied to a field experiment to investigate differences in root growth of acid-soil resistant and sensitive genotypes of perennial pasture grasses in an acid soil. DNA was extracted directly from soil and species-specific DNA was quantified using quantitative real-time PCR prior to estimation of root biomass.

Results

Root growth of the perennial grasses was quantified using the DNA-based technique, although separate calibration procedures were needed to convert DNA content to root mass for each species, soil layer and sampling date. Compared to acid-soil resistant genotypes, lesser root growth in acid soil layers and reduced above-ground dry matter production was observed for acid-soil sensitive genotypes.

Conclusions

The DNA-based method allowed genotypic differences in root growth to be assessed directly in soil and was advantageous for rapid processing of a large number of samples. However, high replication was still required to overcome spatial variability and separate calibrations were required for different species and soil depths across sampling times. The technique demonstrated greater root growth of acid-soil resistant perennial grasses which was beneficial for their establishment and persistence.

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Acknowledgments

This work was funded by a Hackett Postgraduate Research Scholarship awarded to REH (The University of Western Australia), CSIRO and Meat and Livestock Australia. We thank R. Veness, ‘Snowdonia’, Rye Park, for kindly allowing us to conduct this work on his land. Thank you to Adam Stefanski, Phil Armstrong and Scott McDonald for technical assistance. The Molecular Diagnostics Group at the South Australian Research and Development Institute conducted quantitative real-time PCR assays as part of the DNA extraction service. Valuable discussions with their staff are also acknowledged. Alec Zwart provided assistance with statistical analysis.

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Author notes

  1. Rebecca E. Haling

    Present address: School of Environmental and Rural Science, University of New England, Armidale, NSW 2351, Australia

Authors and Affiliations

  1. CSIRO Sustainable Agriculture National Research Flagship/CSIRO Plant Industry, GPO Box 1600, Canberra, ACT 2601, Australia

    Rebecca E. Haling, Richard J. Simpson, Richard A. Culvenor & Alan E. Richardson

  2. School of Plant Biology and Institute of Agriculture, The University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia

    Rebecca E. Haling, Richard J. Simpson, Hans Lambers & Alan E. Richardson

Authors
  1. Rebecca E. Haling
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Correspondence to Alan E. Richardson.

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Responsible Editor: Peter J. Gregory.

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Haling, R.E., Simpson, R.J., Culvenor, R.A. et al. Field application of a DNA-based assay to the measurement of roots of perennial grasses. Plant Soil 358, 183–199 (2012). https://doi.org/10.1007/s11104-012-1405-2

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  • DOI: https://doi.org/10.1007/s11104-012-1405-2

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