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Penicillin Trunk Injection Affects Bacterial Community Structure in Citrus Trees

  • Marina S. Ascunce
  • Keumchul Shin
  • Jose C. Huguet-Tapia
  • Ravin Poudel
  • Karen A. Garrett
  • Ariena H. C. van Bruggen
  • Erica M. Goss
Plant Microbe Interactions

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.

Keywords

Citrus Huanglongbing Microbiomes 16S rRNA Antibiotics Networks 

Notes

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.

Supplementary material

248_2018_1302_MOESM1_ESM.pdf (3.6 mb)
ESM 1 (PDF 3731 kb)

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Authors and Affiliations

  1. 1.Emerging Pathogens InstituteUniversity of FloridaGainesvilleUSA
  2. 2.Department of Plant PathologyUniversity of FloridaGainesvilleUSA
  3. 3.Department of Plant, Soil and Microbial SciencesMichigan State UniversityEast LansingUSA
  4. 4.Institute for Sustainable Food SystemsUniversity of FloridaGainesvilleUSA

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