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Visualisation of the mode of action of a biological control agent, Aureobasidium pullulans (strain YBCA5) against Pseudomonas syringae pv. actinidiae biovar 3 on the kiwifruit phylloplane

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Abstract

The bio-bactericide YBCA5 is a strain of Aureobasidium pullulans (de Bary) G. Arnaud, which mitigates the leaf spot symptoms of bacterial canker in kiwifruit caused by Pseudomonas syringae pv. actinidiae (Psa) biovar 3. Using scanning electron microscopy (SEM) this paper explores the mode of action of YBCA5 against Psa by visualising the interactions between the two microorganisms in planta. Time course studies were conducted by applying the abaxial surface of kiwifruit leaves (Actinidia chinensis var. deliciosa ‘Hayward’) with either YBCA5 or Psa separately and the two microorganisms in combination. SEM revealed that blastospores of YBCA5 occasionally lodged inside the outer cavity of the stomata and commenced multiplying (budding) within 7 days after application. Within 10–14 days, mycelium was observed extending across the phylloplane. On leaves applied with both YBCA5 and Psa, the two microorganisms could be differentiated based on differences in their size with Psa cells measuring approximately a quarter of the length of the YBCA5 blastospores (1.6 ± 0.02 µm and 6.1 ± 0.2 µm, respectively). The Psa cells adhered to the YBCA5 blastospores and hyphae. At 7 days after inoculation, Psa cells adjacent to YBCA5 were deflated with their typical rod-shape distorted and appeared unable to infect the stomata. On average, distorted Psa cells were 20% shorter and 30% wider compared with those on leaves without YBCA5. At 10 days after inoculation, the Psa cells were appressed against the phylloplane and appeared to have collapsed. We hypothesise that this phenomenon is associated with extracellular substances exuded by YBCA5.

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Acknowledgements

We thank Ms Ria Rebstock and Dr Beccy Ganley, Plant & Food Research, for feedback on the manuscript and Ms Lindy Guo, Plant & Food Research, for statistical assistance. Ms Janine Johnson, Plant & Food Research, provided helpful editorial comments. We greatly appreciate the constructive review by an anonymous referee. We are grateful to Dr Maureen O’Callaghan (AgResearch Programme Leader) for supporting this project. This research was funded by the New Zealand Ministry of Business, Innovation and Employment (MBIE) programme ‘Next Generation Biopesticides’ (Contract number C10X1310).

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  1. The New Zealand Institute for Plant and Food Research Limited, Auckland, New Zealand

    P. A. Rheinländer & P. W. Sutherland

  2. The New Zealand Institute for Plant and Food Research Limited, Hamilton, New Zealand

    P. A. G. Elmer

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  1. P. A. Rheinländer
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Conceptualization: P.A.R., P.W.S. and P.A.G.E.; Methodology: P.A.R. and P.W.S.; Formal analysis and investigation: P.W.S. and P.A.R.; Writing-original draft preparation: P.A.R.; Writing-review and editing: P.A.R., P.W.S. and P.A.G.E.; Funding acquisition: P.A.G.E.

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13313_2021_783_MOESM1_ESM.jpg

Supplementary file1. Scanning electron micrographs of fibrillary deposits (arrows) observed associated with Pseudomonas syringae pv. actinidiae biovar 3 (Psa) and YBCA5 (a strain of Aureobasidium pullulans (de Bary) G. Arnaud) on the abaxial phylloplane of Actinidia chinensis var. deliciosa ‘Hayward’. (A) Psa cells on a leaf inoculated with Psa only, (B) Psa cells adhering to a hypha of YBCA5 on a leaf applied with both organisms, (C) hyphae of YBCA5, and (D) blastospore of YBCA5. (JPG 2111 KB)

13313_2021_783_MOESM2_ESM.jpg

Supplementary file2 (A) Scanning electron micrograph (SEM) of the abaxial phylloplane of Actinidia chinensis var. deliciosa ‘Hayward’ 7 days after application with YBCA5 (a strain of Aureobasidium pullulans (de Bary) G. Arnaud) and Pseudomonas syringae pv. actinidiae biovar 3 (Psa). Blastospores of YBCA5 (black arrows) wedged in a crevice of the phylloplane and isolated from Psa cells. In areas with no adjacent YBCA5 the Psa cells (white arrow) were attracted to and able to infect the stomata. (B) SEM of the epidermal cells of the abaxial phylloplane of an untreated control leaf of A. chinensis var. deliciosa ‘Hayward’. No changes of the epidermis were recorded between treated leaves and the untreated controls. (JPG 1036 KB)

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Rheinländer, P.A., Sutherland, P.W. & Elmer, P.A.G. Visualisation of the mode of action of a biological control agent, Aureobasidium pullulans (strain YBCA5) against Pseudomonas syringae pv. actinidiae biovar 3 on the kiwifruit phylloplane. Australasian Plant Pathol. 50, 379–388 (2021). https://doi.org/10.1007/s13313-021-00783-3

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