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Penicillin Trunk Injection Affects Bacterial Community Structure in Citrus Trees

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Abstract

Huanglongbing (HLB), caused by Candidatus Liberibacter asiaticus (CLas), an uncultured α-proteobacterium, is the most destructive disease of citrus trees worldwide. In previous studies, trunk injections of penicillin reduced CLas titers and HLB symptoms in citrus. However, antibiotic effects on the whole plant microbial community, which include effects on taxa that interact with CLas, have not yet been addressed. In this study, we investigated the effects of penicillin injection (0, 1000, and 6000 mg L−1) on rhizospheric and endophytic bacterial communities of grapefruit trees in field and greenhouse experiments through culture-independent high-throughput sequencing. DNA extractions from petioles and roots were subjected to 16S rRNA high-throughput sequencing, and reads were clustered by sequence similarity into operational taxonomic units (OTUs). Principal coordinates analysis based on weighted-UniFrac distances did not reveal differences in bacterial communities among treatments in any of the sample sources. However, pairwise linear discriminant analysis indicated significant differences in relative abundance of some taxa (including CLas) among treatments. Network analysis showed that penicillin produced major changes in root bacterial community structure by affecting interspecific microbial associations. This study provides new knowledge of the effect of antimicrobial treatments on interspecific relationships in citrus microbial communities.

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Acknowledgments

This research was funded by the Florida Department of Agriculture and Consumer services; additional funding was provided by the University of Florida (UF) IFAS Research Early Career Seed Fund, the UF Emerging Pathogens Institute, the UF Department of Plant Pathology, and the Esther B. O’Keeffe Foundation. We are grateful to Robert H. Moore of the Pesticide Registration Review Section, Bureau of Scientific Evaluation and Technical Assistance, Division of Agricultural Environmental Services, Florida Department of Agriculture and Consumer Services, Tallahassee, FL, for providing the exemption to use penicillin in an experimental field, and in a BSL2 greenhouse and laboratory. Hossein A. Narouei-Khandan, Oluwaseun Olawale Kolawole, Cody Neff, Hannah Fahsbender, Ellen Dickstein, and Yorley Nikary Bustamante participated in collecting and processing citrus samples. Mark G. Kann and Jim A. Boyer took care of the young citrus trees in the greenhouse at Citra. We wish to thank the UF Plant Pathology Department for providing support for greenhouse experiments, especially Mike Stilwell and his team. We are also grateful to the High Performance Computing Center and the HiPerGator cluster support staff for their assistance. We want to thank the editor and the reviewers for comments on an earlier version of the manuscript.

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  1. Keumchul Shin

    Present address: Department of Plant, Soil and Microbial Sciences, Michigan State University, East Lansing, MI, USA

Authors and Affiliations

  1. Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA

    Marina S. Ascunce, Keumchul Shin, Ravin Poudel, Karen A. Garrett, Ariena H. C. van Bruggen & Erica M. Goss

  2. Department of Plant Pathology, University of Florida, Gainesville, FL, USA

    Marina S. Ascunce, Keumchul Shin, Jose C. Huguet-Tapia, Ravin Poudel, Karen A. Garrett, Ariena H. C. van Bruggen & Erica M. Goss

  3. Institute for Sustainable Food Systems, University of Florida, Gainesville, FL, USA

    Ravin Poudel & Karen A. Garrett

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  1. Marina S. Ascunce
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Correspondence to Marina S. Ascunce.

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Ascunce, M.S., Shin, K., Huguet-Tapia, J.C. et al. Penicillin Trunk Injection Affects Bacterial Community Structure in Citrus Trees. Microb Ecol 78, 457–469 (2019). https://doi.org/10.1007/s00248-018-1302-1

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