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RESEARCH ARTICLE (Open Access)
Contents Vol 74(12)

Screening and field evaluation of white clover rhizobia for New Zealand pastures

Shengjing Shi https://orcid.org/0000-0002-8938-1932 A * , Steve Wakelin B , Emily Gerard A , Sandra Young A , Chikako van Koten A , John Caradus C , Andrew G. Griffiths https://orcid.org/0000-0002-0573-1668 D , Ross A. Ballard https://orcid.org/0000-0003-0814-3586 E and Maureen O’Callaghan A
+ Author Affiliations
- Author Affiliations

A AgResearch Ltd, Lincoln Science Centre, Christchurch, New Zealand.

B Scion, PO Box 29237, Christchurch 8440, Christchurch, New Zealand.

C Grasslanz Technology Ltd., Hamilton, New Zealand.

D AgResearch Ltd, Grasslands Research Centre, Private Bag 11008, Palmerston North 4410, New Zealand.

E South Australian Research and Development Institute, GPO Box 397, Adelaide, SA 5001, Australia.

* Correspondence to: shengjing.shi@agresearch.co.nz

Handling Editor: Matthew Denton

Crop & Pasture Science 74(12) 1258-1271 https://doi.org/10.1071/CP22405
Submitted: 20 December 2022  Accepted: 30 May 2023  Published: 20 June 2023

© 2023 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC)

Abstract

Context

Biological nitrogen (N) fixation through the rhizobia–legume symbiosis is a sustainable and cost-effective source of N for agriculture. In New Zealand (NZ), white clover (Trifolium repens) is a key component of pastures and rhizobial inoculation of clover is widely used. The current commercial inoculant for white clover, TA1, was isolated in Australia in the 1950s and may not be the best partner for modern white clover cultivars.

Aims

To identify Rhizobium leguminosarum bv. trifolii (Rlt) isolates suitable for use in NZ pastures.

Methods

The symbiotic potential of >230 isolates collected from throughout NZ was evaluated in plant bioassays. Selected isolates were further evaluated in pot and field trials.

Key results

Approximately 40% of NZ isolates supported better clover growth than TA1 under N-limited conditions in vitro. Of 24 Rlt isolates evaluated in a glasshouse trial, five produced significantly higher clover biomass than TA1. Three (S11N9, S20N7, S4N6) of nine isolates evaluated in two field trials in 2018–2019 significantly increased clover growth (12–38%) compared with paired uninoculated clover at several harvests, whereas inoculation with TA1 did not improve yield. In a third trial in 2020, S11N9 and S20N7 increased clover growth compared with the uninoculated control at two of three harvests; S4N6 performed better than TA1 at one harvest. When tested with four white clover cultivars, five Rlt isolates had higher symbiotic potential than TA1.

Conclusions

Inoculating white clover would be beneficial if improved inoculant isolates were available.

Implications

We recommend some NZ Rlt isolates could be developed into commercial inoculants to improve white clover performance in NZ.

Keywords: biological N fixation, cultivars, field trials, inoculant, Rhizobium leguminosarum bv. trifolii, sustainability, symbiotic potential, white clover.

References

Abdel-Salam MS, Ibrahim SA, Adb-El-Halim MM, Badawy FM, Abo-Aba SEM (2010) Phenotypic characterization of indigenous Egyptian Rhizobial strains for abiotic stresses performance. Journal of American Science 6, 498-503.
| Google Scholar |

Bashan Y, de-Bashan LE, Prabhu SR, Hernandez J-P (2014) Advances in plant growth-promoting bacterial inoculant technology: formulations and practical perspectives (1998–2013). Plant and Soil 378, 1-33.
| Crossref | Google Scholar |

Boddey RM, Peoples MB, Palmer B, Dart PJ (2000) Use of the 15N natural abundance technique to quantify biological nitrogen fixation by woody perennials. Nutrient Cycling in Agroecosystems 57, 235-270.
| Crossref | Google Scholar |

Bonish PM (1980) Nodulation of white clover: plant influences on the effectiveness of Rhizobium trifolii. New Zealand Journal of Agricultural Research 23, 239-242.
| Crossref | Google Scholar |

Braakhekke MC, Rebel KT, Dekker SC, Smith B, Beusen AHW, Wassen MJ (2017) Nitrogen leaching from natural ecosystems under global change: a modelling study. Earth System Dynamics 8, 1121-1139.
| Crossref | Google Scholar |

Brockwell J (1980) Experiments with crop and pasture legumes – priciples and practice. In ‘Methods for evaluationg biological nitrogen fixation’. (Ed. F Bergerson) pp. 417–488. (Wiley: Brisbane, Qld, Australia)

Brockwell J, Holliday RA, Pilka A (1988) Evaluation of the symbiotic nitrogen-fixing potential of soils by direct microbiological means. Plant and Soil 108, 163-170.
| Crossref | Google Scholar |

Bullard GK, Roughley RJ, Pulsford DJ (2005) The legume inoculant industry and inoculant quality control in Australia: 1953-2003. Australian Journal of Experimental Agriculture 45, 127-140.
| Crossref | Google Scholar |

Caradus JR, Hay MJM, Mackay AD, Thomas VJ, Dunlop J, Lambert MG, Hart AL, van den Bosch J, Wewala S (1993) Variation within white clover (Trifolium repens L.) for phenotypic plasticity of morphological and yield related characters, induced by phosphorus supply. New Phytologist 123, 175-184.
| Crossref | Google Scholar |

Caradus JR, Hay RJM, Woodfield DR (1995) The positioning of white clover cultivars in New Zealand. In ‘White clover: New Zealand’s competitive edge. Vol. 6’. (Ed. D Woodfield) pp. 45–49. (New Zealand Grassland Association)

Caradus JR, Woodfield DR, Stewart AV (1996) Overview and vision for white clover. In ‘White clover: New Zealand’s competitive edge. Agronomy Society of New Zealand Special Publication No 11/Grassland Research and Practice Series. Vol. 6’. pp. 1–6. (New Zealand Grassland Association)

Caradus JR, Clifford PTP, Chapman DF, Cousins GR, Williams WM, Miller JE (1997) Breeding and description of ‘Grasslands Sustain’, a medium-large leaved white clover (Trifolium repens L.) cultivar. New Zealand Journal of Agricultural Research 40, 1-7.
| Crossref | Google Scholar |

Cooper BM, Chapman DF (1993) Grasslands Prestige (G.39), a white clover cultivar originating from northern New Zealand. In ‘Proceedings of International Grassland Congress. Vol. 17’. pp. 458–459. (New Zealand Grassland Association)

Deaker R, Roughley RJ, Kennedy IR (2004) Legume seed inoculation technology – a review. Soil Biology and Biochemistry 36, 1275-1288.
| Crossref | Google Scholar |

Deaker R, Hartley E, Gemell G (2012) Conditions affecting shelf-life of inoculated legume seed. Agriculture 2, 38-51.
| Crossref | Google Scholar |

Delestre C, Laugraud A, Ridgway H, Ronson C, O’Callaghan M, Barrett B, Ballard R, Griffiths A, Young S, Blond C, Gerard E, Wakelin S (2015) Genome sequence of the clover symbiont Rhizobium leguminosarum bv. trifolii strain CC275e. Standards in Genomic Sciences 10, 121.
| Crossref | Google Scholar |

Drew EA, Charman N, Dingemanse R, Hall E, Ballard RA (2011) Symbiotic performance of Mediterranean Trifolium spp. with naturalised soil rhizobia. Crop & Pasture Science 62, 903-913.
| Crossref | Google Scholar |

Evans J, Hochman Z, O’Connor GE, Osborne GJ (1988) Soil acidity and Rhizobium: their effects on nodulation of subterranean clover on the slopes of southern New South Wales. Australian Journal of Agricultural Research 39, 605-618.
| Crossref | Google Scholar |

Ferguson S, Major AS, Sullivan JT, Bourke SD, Kelly SJ, Perry BJ, Ronson CW, Vieille C (2020) Rhizobium leguminosarum bv. trifolii NodD2 enhances competitive nodule colonization in the clover-rhizobium symbiosis. Applied and Environmental Microbiology 86, e01268-20.
| Crossref | Google Scholar |

Ford JL, Cousins GR, Jahufer Z, Baird IJ, Woodfield DR, Barrett BA (2015) Grasslands legacy – a new, large-leaved white clover cultivar with broad adaption. Journal of New Zealand Grasslands 77, 211-218.
| Crossref | Google Scholar |

Gibson AH, Date RA, Ireland JA, Brockwell J (1976) A comparison of competitiveness and persistence amongst five strains of Rhizobium tripolii. Soil Biology and Biochemistry 8, 395-401.
| Crossref | Google Scholar |

Graham PH (2008) Ecology of the root-nodule bacteria of legumes. In ‘Nitrogen-fixing leguminous symbioses’. (Eds MJ Dilworth, EK James, JI Sprent, WE Newton) pp. 23–58. (Springer: Berlin, Germany)

Greenwood RM (1965) Populations of rhizobia in New Zealand soils. Proceedings of the New Zealand Grasslands Association 26, 95-101.
| Google Scholar |

Griffiths AG, Moraga R, Tausen M, Gupta V, Bilton TP, Campbell MA, Ashby R, Nagy I, Khan A, Larking A, Anderson C, Franzmayr B, Hancock K, Scott A, Ellison NW, Cox MP, Asp T, Mailund T, Schierup MH, Andersen SU (2019) Breaking free: the genomics of allopolyploidy-facilitated niche expansion in white clover. The Plant Cell 31, 1466-1487.
| Crossref | Google Scholar |

Harrison SP, Young JPW, Jones DG (1989) Rhizobium population genetics: host preference and strain competition effects on the range of Rhizobium leguminosarum biovar Trifolii genotypes isolated from natural populations. Soil Biology and Biochemistry 21, 981-986.
| Crossref | Google Scholar |

Hayes RC, Rohan M, Li GD, Orgill SE, Poile GJ, Oates AA, Conyers MK (2022) The nature of spatial variability of four soil chemical properties and the implications for soil sampling. Journal of Soils and Sediments 22, 3006-3017.
| Crossref | Google Scholar |

Högberg P (1997) Tansley review no. 95 15N natural abundance in soil-plant systems. New Phytologist 137, 179-203.
| Crossref | Google Scholar |

Howieson JG, Yates RJ, O’Hara GW, Ryder M, Real D (2005) The interactions of Rhizobium leguminosarum biovar trifolii in nodulation of annual and perennial Trifolium spp. from diverse centres of origin. Australian Journal of Experimental Agriculture 45, 199-207.
| Crossref | Google Scholar |

Irisarri P, Cardozo G, Tartaglia C, Reyno R, Gutiérrez P, Lattanzi FA, Rebuffo M, Monza J (2019) Selection of competitive and efficient rhizobia strains for white clover. Frontiers in Microbiology 10, 768.
| Crossref | Google Scholar |

Jahufer MZZ, Dunn A, Baird I, Ford JL, Griffiths AG, Jones CS, Woodfield DR, Barrett BA (2013) Genotypic variation for morphological traits in a white clover mapping population evaluated across two environments and three years. Crop Science 53, 460-472.
| Crossref | Google Scholar |

Jahufer MZZ, Ford JL, Cousins GR, Woodfield DR (2021) Relative performance of white clover (Trifolium repens) cultivars and experimental synthetics under rotational grazing by beef cattle, dairy cattle and sheep. Crop & Pasture Science 72, 926-938.
| Crossref | Google Scholar |

Lowther WL, Kerr GA (2011) White clover seed inoculation and coating in New Zealand. Proceedings of the New Zealand Grassland Association 73, 93-102.
| Crossref | Google Scholar |

McNeil DL (1982) Variations in ability of Rhizobium japonicum strains to nodulate soybeans and maintain fixation in the presence of nitrate. Applied and Environmental Microbiology 44, 647-652.
| Crossref | Google Scholar |

Melino VJ, Drew EA, Ballard RA, Reeve WG, Thomson G, White RG, O’Hara GW (2012) Identifying abnormalities in symbiotic development between Trifolium spp. and Rhizobium leguminosarum bv. trifolii leading to sub-optimal and ineffective nodule phenotypes. Annals of Botany 110, 1559-1572.
| Crossref | Google Scholar |

Moeskjær S, Skovbjerg CK, Tausen M, Wind R, Roulund N, Janss L, Andersen SU (2022) Major effect loci for plant size before onset of nitrogen fixation allow accurate prediction of yield in white clover. Theoretical and Applied Genetics 135, 125-143.
| Crossref | Google Scholar |

Nguyen HP, Miwa H, Obirih-Opareh J, Suzaki T, Yasuda M, Okazaki S (2020) Novel rhizobia exhibit superior nodulation and biological nitrogen fixation even under high nitrate concentrations. FEMS Microbiology Ecology 96, fiz184.
| Crossref | Google Scholar |

Olsson L, Rugbjerg P, Torello Pianale L, Trivellin C (2022) Robustness: linking strain design to viable bioprocesses. Trends in Biotechnology 40, 918-931.
| Crossref | Google Scholar |

Onishchuk OP, Vorobyov NI, Provorov NA (2017) Nodulation competitiveness of nodule bacteria: genetic control and adaptive significance: review. Applied Biochemistry and Microbiology 53, 131-139.
| Crossref | Google Scholar |

Rigg JL, Webster AT, Harvey DM, Orgill SE, Galea F, Dando AG, Collins DP, Harris CA, Newell MT, Badgery WB, Hayes RC (2021) Cross-host compatibility of commercial rhizobial strains for new and existing pasture legume cultivars in south-eastern Australia. Crop & Pasture Science 72, 652-665.
| Crossref | Google Scholar |

Rys GJ, Bonish PM (1981) Effectiveness of Rhizobium trifolii populations associated with Trifolium species in Taranaki, New Zealand. New Zealand Journal of Experimental Agriculture 9, 329-335.
| Crossref | Google Scholar |

Sessitsch A, Pfaffenbichler N, Mitter B (2019) Microbiome applications from lab to field: facing complexity. Trends in Plant Science 24, 194-198.
| Crossref | Google Scholar |

Sherwood MT, Masterson CL (1974) Importance of using the correct test host in assessing the effectiveness of indigenous populations of Rhizobium Trifolii. Irish Journal of Agricultural Research 13, 101-108.
| Google Scholar |

Shi S, Villamizar L, Gerard E, Ronson C, Wakelin S, Ballard R, Caradus JR, O’Callaghan M (2019) Increasing biological nitrogen fixation by white clover-rhizobia symbiosis. Journal of New Zealand Grasslands 81, 231-234.
| Crossref | Google Scholar |

Slattery JF, Coventry DR, Slattery WJ (2001) Rhizobial ecology as affected by the soil environment. Australian Journal of Experimental Agriculture 41, 289-298.
| Crossref | Google Scholar |

Soumare A, Diedhiou AG, Thuita M, Hafidi M, Ouhdouch Y, Gopalakrishnan S, Kouisni L (2020) Exploiting biological nitrogen fixation: a route towards a sustainable agriculture. Plants 9, 1011.
| Crossref | Google Scholar |

StatsNZ (2021) Fertilisers – nitrogen and phosphorus. Available at https://www.stats.govt.nz/indicators/fertilisers-nitrogen-and-phosphorus

Trivellin C, Olsson L, Rugbjerg P (2022) Quantification of microbial robustness in yeast. ACS Synthetic Biology 11, 1686-1691.
| Crossref | Google Scholar |

Unkovich M, Herridge D, Peoples M, Cadisch G, Boddey R, Giller K, Alves B, Chalk P (2008) ‘Measuring plant-associated nitrogen fixation in agricultural systems.’ ACIAR Monograph 136. (Australian Centre for International Agricultural Research)

van Ham R, O’Callaghan M, Geurts R, Ridgway HJ, Ballard R, Noble A, Macara G, Wakelin SA (2016) Soil moisture deficit selects for desiccation tolerant Rhizobium leguminosarum bv. trifolii. Applied Soil Ecology 108, 371-380.
| Crossref | Google Scholar |

Wakelin S, Tillard G, van Ham R, Ballard R, Farquharson E, Gerard E, Geurts R, Brown M, Ridgway H, O’Callaghan M (2018) High spatial variation in population size and symbiotic performance of Rhizobium leguminosarum bv. trifolii with white clover in New Zealand pasture soils. PLoS ONE 13, e0192607.
| Crossref | Google Scholar |

Weith SK, Jahufer MZZ, Hofmann RW, Anderson CB, Luo D, Ehoche OG, Cousins G, Jones EE, Ballard RA, Griffiths AG (2022) Quantitative genetic analysis reveals potential to breed for improved white clover growth in symbiosis with nitrogen-fixing Rhizobium bacteria. Frontiers in Plant Science 13, 953400.
| Crossref | Google Scholar |

Woodfield DR, Clifford PTP, Cousins GR, Ford JL, Baird IJ, Miller JE, Woodward SL, Caradus JR (2001) Grasslands Kopu II and Crusader: new generation white clovers. Proceedings of the New Zealand Grasslands Association 63, 103-108.
| Crossref | Google Scholar |

Woodfield DR, Clifford PTP, Baird IJ, Cousins GR, Miller JE, Widdup KH, Caradus JR (2003) Grasslands tribute: a multi-purpose white clover for Australasia. Proceedings of the New Zealand Grassland Association 65, 157-162.
| Crossref | Google Scholar |

Zahran HH (1999) Rhizobium-legume symbiosis and nitrogen fixation under severe conditions and in an arid climate. Microbiology and Molecular Biology Reviews 63, 968-989.
| Crossref | Google Scholar |