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An efficient petiole-feeding bioassay for introducing aqueous solutions into dicotyledonous plants

Nature Protocols volume 6, pages 36–45 (2011)Cite this article

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Abstract

Introducing bioactive molecules into plants helps establish their roles in plant growth and development. Here we describe a simple and effective petiole-feeding protocol to introduce aqueous solutions into the vascular stream and apoplast of dicotyledonous plants. This 'intravenous feeding' procedure has wide applicability to plant physiology, specifically with regard to the analysis of source-sink allocations, long-distance signaling, hormone biology and overall plant development. In comparison with existing methods, this technique allows the continuous feeding of aqueous solutions into plants without the need for constant monitoring. Findings are provided from experiments using soybean plants fed with a range of aqueous solutions containing tracer dyes, small metabolites, radiolabeled chemicals and biologically active plant extracts controlling nodulation. Typically, feeding experiments consist of (i) generating samples to feed (extracts, solutions and so on); (ii) growing recipient plants; (iii) setting up the feeding apparatus; and (iv) feeding sample solutions into the recipient plants. When the plants are ready, the feeding procedure can take 1–3 h to set up depending on the size of experiment (not including preparation of materials). The petiole-feeding technique also works with other plant species, including tomato, chili pepper and cabbage plants, as demonstrated here.

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Figure 1: Construction of the feeding assembly.
Figure 2: Attachment of the feeding assembly to a recipient soybean plant.
Figure 3: Translocation of radioactive mannitol following petiole feeding in soybean (Bragg) plants.
Figure 4: Visualization of brilliant blue dye.
Figure 5: Suppression of nodule development in soybean.

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Acknowledgements

We thank the Australian Research Council for provision of a Centre of Excellence grant (CEO348212); we also thank the Queensland State Government Smart State Innovation Scheme and the University of Queensland Strategic Fund for support. We thank D. Reid (Centre for Integrative Legume Research) for the RMH formulation and C. Atkins (University of Western Australia) for providing lupin phloem.

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Authors and Affiliations

  1. Australian Research Council Centre of Excellence for Integrative Legume Research, The University of Queensland, Brisbane, Queensland, Australia

    Yu-Hsiang Lin, Meng-Han Lin, Peter M Gresshoff & Brett J Ferguson

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  1. Yu-Hsiang Lin
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  2. Meng-Han Lin
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Contributions

All authors contributed equally to the design of the technique and to the composition of the paper. Y.-H.L. and M.-H.L. were responsible for the development and optimization of the experimental technique. Y.-H.L. was responsible for data acquisition and analysis. P.M.G. and B.J.F. provided critical creative input and supervision for the development of the technique.

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Correspondence to Peter M Gresshoff.

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Lin, YH., Lin, MH., Gresshoff, P. et al. An efficient petiole-feeding bioassay for introducing aqueous solutions into dicotyledonous plants. Nat Protoc 6, 36–45 (2011). https://doi.org/10.1038/nprot.2010.171

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