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The free-living rhizosphere fungus Trichoderma hamatum GD12 enhances clover productivity in clover-ryegrass mixtures

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Abstract

Aim

A principal goal of grassland management is to minimize the use of artificial fertilizers by maximising the productivity of nitrogen-fixing leguminous plants such as clovers. The objective of this study was to investigate whether a plant-growth-promoting strain of the free-living rhizosphere fungus Trichoderma hamatum (GD12) could be used as a natural and sustainable means of enhancing the competitiveness of white clover (Trifolium repens) while allowing increased productivity of both clover and ryegrass (Lolium perenne) in mixed species systems.

Methods

An assay was conducted in rhizotrons with white clover and ryegrass sown alone and in mixture and in soils inoculated and non-inoculated with GD12. Plant height, growing rate, phenological stage, number of Rhizobium nodules and biomass were assessed. A histological study of Rhizobium nodules and a stable isotopes analysis was conducted to determine the N fixation capacity of white clover.

Results

When introduced as a soil inoculant, the fungus increased biomass production of both plant species and shortened their phenological cycles. Furthermore, in clover, GD12 enhanced plant height and growth rate and stimulated Rhizobium nodulation, while 15 N stable isotope analysis demonstrated increased N2 fixation.

Conclusion

This shows that soil amendment with a beneficial strain of saprotrophic fungus bestows a competitive advantage to white clover in clover-ryegrass mixtures and provides a sustainable mechanism for improving the mixture productivity.

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Acknowledgments

The authors would like acknowledge to the Operative Program of European Social Fund (ESF) 2007–2013 “Andalucía se mueve con Europa” and to the project INIA-RTA2011-00031 that funded the contract and stay of C. Alcántara in North Wyke Rothamsted Research. Rothamsted Research is supported by the UK Biotechnology and Biological Sciences Research Council (BBSRC). The authors would to thank Dr. Domínguez for reviewing the statistical methods used in this study.

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Authors and Affiliations

  1. Área de Producción Agraria, IFAPA Centro Alameda del Obispo, Junta de Andalucía, Avda. Menéndez Pidal s/n, 14080, Córdoba, Spain

    C. Alcántara & V. Pedraza

  2. Área de Mejora y Biotecnología, IFAPA Centro Alameda del Obispo, Junta de Andalucía, Avda. Menéndez Pidal s/n, 14080, Córdoba, Spain

    A. Pérez-de-Luque

  3. Biosciences, University of Exeter, Geoffrey Pope Building, Stocker Road, Exeter, EX4 4QD, UK

    C. R. Thornton

  4. Sustainable Soils and Grassland Systems Department, Rothamsted Research, North Wyke, Okehampton, EX20 2SB, UK

    K. Le Cocq & P. J. Murray

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  1. C. Alcántara
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  2. C. R. Thornton
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  3. A. Pérez-de-Luque
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  4. K. Le Cocq
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  5. V. Pedraza
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Corresponding author

Correspondence to C. Alcántara.

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Responsible Editor: Duncan D. Cameron.

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Fig. 1

Cross sections of Rhizobium nodules on roots of white clover. a) General view of a nodule from clover growing in un-inoculated soil. b) Detail of a (square I) showing cells with bacteroids at the infection zone. c) Detail of a (square II) showing infected cells at the nitrogen-fixing zone. d) Detail of a (square III) showing infected cells at the senescence zone. e) Detailed picture of cells from the nitrogen-fixing zone showing bacteroids with rod-shaped form. f) General view of a nodule from clover growing in Trichoderma inoculated soil. g) Detail of f (square I) showing cells with bacteroids at the infection zone. h) Detail of f (square II) showing infected cells at the nitrogen-fixing zone. Notice the presence of cells containing starch and amyloplasts (arrows). i) Detail of f (square III) showing infected cells at the senescence zone. j) Detailed picture of cells from the nitrogen-fixing zone showing bacteroids with rod-shaped form and the presence of amyloplasts inside some cells (arrows) (TIFF 29214 kb)

High Resolution (GIF 14269 kb)

Fig. 2

Rhizotrons from two replicates of the different species: clover (a, b), ryegrass (c, d) and mixture (e, f); and treatments (with or without T). Notice the differences in plant size between rhizotrons inoculated with GD12 and those un-inoculated (TIFF 567 kb)

High Resolution (GIF 294 kb)

Fig. 3

Root morphologies of clover and ryegrass plants grown in soil inoculated with T. hamatum GD12 (+T) or in un-inoculated soil (−T). Scale bar = 7,5 cm. (TIFF 188 kb)

High Resolution (GIF 10 kb)

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Alcántara, C., Thornton, C.R., Pérez-de-Luque, A. et al. The free-living rhizosphere fungus Trichoderma hamatum GD12 enhances clover productivity in clover-ryegrass mixtures. Plant Soil 398, 165–180 (2016). https://doi.org/10.1007/s11104-015-2646-7

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