Abstract
Trunk injection of antibiotics has re-emerged as a strategy to mitigate citrus huanglongbing (HLB), a devastating disease associated with the bacterium Candidatus Liberibacter asiaticus (CLas). Despite commercial adoption in Florida, no studies have examined how oxytetracycline (OTC) injection may impact active plant-associated microbial communities. We investigated the impact of OTC trunk injection on tree physiological parameters, HLB control, and the prokaryotic and eukaryotic microbiome of the active inner bark and rhizosphere in a commercial citrus orchard under HLB-endemic conditions. OTC injection significantly increased juice quality and fruit weight. OTC was detected in leaves, bark, and roots but concentrations varied with time and across plant tissues. OTC injection reduced the abundance and diversity of the active prokaryotic microbiome in the bark and rhizosphere during the first three months post-injection. Specific prokaryotic taxa were responsive to OTC injection and their relative abundances related to increased plant performance. This study shows that OTC injection can promote the recovery of tree physiological functions impacted by HLB while also maximizing the benefits from certain prokaryotes of the bark and rhizosphere microbiomes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All sequence data for this study have been deposited at NCBI under BioProject PRJNA1048447.
Code Availability
Not applicable.
References
Aćimović SG, Zeng Q, McGhee GC, Sundin GW, Wise JC (2015) Control of fire blight (Erwinia amylovora) on apple trees with trunk-injected plant resistance inducers and antibiotics and assessment of induction of pathogenesis-related protein genes. Front Plant Sci 6:123631. https://doi.org/10.3389/fpls.2015.00016
Albrecht U, Bowman KD (2008) Gene expression in Citrus sinensis (L.) Osbeck following infection with the bacterial pathogen Candidatus Liberibacter asiaticus causing Huanglongbing in Florida. Plant Sci 175:291–306. https://doi.org/10.1016/j.plantsci.2008.05.001
Alvarenga DO, Franco MW, Sivonen K, Fiore MF, Varani AM (2020) Evaluating Eucalyptus leaf colonization by Brasilonema Octagenarum (Cyanobacteria, Scytonemataceae) using in planta experiments and genomics. PeerJ 8:e9158. https://nam10.safelinks.protection.outlook.com/?url=https%3A%2F%2Fdoi.org%2F10.7717%2Fpeerj.9158&data=05%7C02%7Cstrauss%40ufl.edu%7Ce2a89a485f48455bbe6508dc57ee8b2c%7C0d4da0f84a314d76ace60a62331e1b84%7C0%7C0%7C638481929250213816%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C0%7C%7C%7C&sdata=JkfUGG9eTeaSUAIScyf1UagCFw5SVnk8NJp77ewOsk8%3D&reserved=0
Amaral-Zettler LA, McCliment EA, Ducklow HW, Huse SM (2009) A method for studying protistan diversity using massively parallel sequencing of V9 hypervariable regions of small-subunit ribosomal RNA genes. PLoS ONE 4:e6372. https://doi.org/10.1371/journal.pone.0006372
Archer L, Albrecht U (2023) Evaluation of trunk injection techniques for systemic delivery of huanglongbing therapies in citrus. HortScience 58:768–778. https://doi.org/10.21273/HORTSCI17172-23
Archer L, Crane JH, Albrecht U (2022a) Trunk injection as a tool to deliver plant protection materials—an overview of basic principles and practical considerations. Horticulturae 8:552. https://doi.org/10.3390/horticulturae8060552
Archer L, Kunwar S, Alferez F, Batuman O, Albrecht U (2022b) Trunk injection of oxytetracycline for huanglongbing management in mature grapefruit and sweet orange trees. Phytopathology 113:1010–1021. https://doi.org/10.1094/phyto-09-22-0330-r
Archer L, Qureshi J, Albrecht U (2022c) Efficacy of trunk injected imidacloprid and oxytetracycline in managing huanglongbing and Asian citrus psyllid in infected sweet orange (Citrus sinensis) trees. Agriculture 12:1592. https://doi.org/10.3390/agriculture12101592
Arrigoni E, Antonielli L, Pindo M, Pertot I, Perazzolli M (2018) Tissue age and plant genotype affect the microbiota of apple and pear bark. Microbiol Res 211:57–68. https://doi.org/10.1016/j.micres.2018.04.002
Arrigoni E, Albanese D, Longa CMO, Angeli D, Donati C, Ioriatti C, Pertot I, Perazzolli M (2020) Tissue age, orchard location and disease management influence the composition of fungal and bacterial communities present on the bark of apple trees. Environ Microbiol 22:2080–2093. https://doi.org/10.1111/1462-2920.14963
Ascunce MS, Shin K, Huguet-Tapia JC, Poudel R, Garrett KA, van Bruggen AHC, Goss EM (2019) Penicillin trunk injection affects bacterial community structure in citrus trees. Microb Ecol 78:457–469. https://doi.org/10.1007/s00248-018-1302-1
Baldwin E, Plotto A, Manthey J, McCollim G, Bai J, Irey M, Cameron R, Luzio G (2010) Effect of Liberibacter infection (huanglongbing disease) of citrus on orange fruit physiology and fruit/fruit juice quality: chemical and physical analyses. J Agric Food Chem 58:1247–1262. https://doi.org/10.1021/jf9031958
Batuman O, Levy A, Brlansky RH (2023) 2023–2024 Florida citrus production guide: blight. https://edis.ifas.ufl.edu/publication/CG038. Accessed 15 November 2023
Bennett JA, Klironomos J (2019) Mechanisms of plant-soil feedback: interactions among biotic and abiotic drivers. New Phytol 222:91–96. https://doi.org/10.1111/nph.15603
Berger C, Laurent F (2019) Trunk injection of plant protection products to protect trees from pests and diseases. Crop prot 124:104831. https://doi.org/10.1016/j.cropro.2019.05.025
Bokulich NA, Kaehler BD, Rideout JR, Dillon M, Bolyen E, Knight R, Huttley GA, Gregory Caporaso J (2018) Optimizing taxonomic classification of marker-gene amplicon sequences with QIIME 2’s q2-feature-classifier plugin. Microbiome 6:1–17. https://doi.org/10.1186/s40168-018-0470-z
Bové JM (2006) Huanglongbing: a destructive, newly-emerging, century-old disease of citrus. J Plant Pathol 88:7–37
Bowman KD, Albrecht U (2020) Rootstock influences on health and growth following Candidatus liberibacter asiaticus infection in young sweet orange trees. Agronomy 10:1907. https://doi.org/10.3390/agronomy10121907
Bulgarelli D, Schlaeppi K, Spaepen S, Van Themaat EVL, Schulze-Lefert P (2013) Structure and functions of the bacterial microbiota of plants. Annu Rev Plant Biol 64:807–838. https://doi.org/10.1146/annurev-arplant-050312-120106
Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP (2016) DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods 13:581–583. https://doi.org/10.1038/nmeth.3869
Carini P, Marsden PJ, Leff JW, Morgan EE, Strickland MS, Fierer N (2016) Relic DNA is abundant in soil and obscures estimates of soil microbial diversity. Nat Microbiol 2:1–6. https://doi.org/10.1038/nmicrobiol.2016.242
Castellano-Hinojosa A, Meyering B, Nuzzo A, Strauss SL, Albrecht U (2021) Effect of plant biostimulants on root and plant health and the rhizosphere microbiome of citrus trees in huanglongbing-endemic conditions. Trees - Struct Funct 35:1525–1539. https://doi.org/10.1007/s00468-021-02133-8
Castellano-Hinojosa A, Albrecht U, Strauss SL (2023a) Interactions between rootstocks and compost influence the active rhizosphere bacterial communities in citrus. Microbiome 11:1–16. https://doi.org/10.1186/s40168-023-01524-y
Castellano-Hinojosa A, Kanissery R, Strauss SL (2023b) Cover crops in citrus orchards impact soil nutrient cycling and the soil microbiome after three years but effects are site-specific. Biol Fertil Soils 59:659–678. https://doi.org/10.1007/s00374-023-01729-1
Cowles RS, Montgomery ME, Cheah CAS-J (2006) Activity and residues of imidacloprid applied to soil and tree trunks to control hemlock woolly adelgid (Hemiptera: Adelgidae) in forests. J Econ Entomol 99:1258–1267. https://doi.org/10.1603/0022-0493-99.4.1258
Crane JH, Wasielweski J, Carrillo D, Gazis R, Schaffer B, Ballen F, Evans E (2020) Recommendations for the detection and mitigation of laurel wilt disease in avocado and related tree species in the home landscape. https://edis.ifas.ufl.edu/publication/HS1358. Accessed 15 November 2023
da Graça JV, Douhan GW, Halbert SE, Keremane ML, Lee RF, Vidalakis G, Zhao H (2016) Huanglongbing: an overview of a complex pathosystem ravaging the world’s citrus. J Integr Plant Biol 58:373–387. https://doi.org/10.1111/jipb.12437
Dahal RH, Chaudhary DK, Kim DU, Kim J (2021) Azohydromonas Caseinilytica sp. nov., a nitrogen-fixing bacterium isolated from forest soil by using optimized culture method. Front Microbiol 12:647132. https://doi.org/10.3389/fmicb.2021.647132
Danilova N, Galitskaya P, Selivanovskaya S (2020) Veterinary antibiotic oxytetracycline’s effect on the soil microbial community. J Ecol Environ 44:1–9. https://doi.org/10.1186/s41610-020-00154-x
Emerson JB, Adams RI, Román CMB, Brooks B, Coil DA, Dahlhausen K, Ganz HH, Hartmann EM, Hsu T, Justice NB, Paulino-Lima IG, Luongo JC, Lymperopoulou DS, Gomez-Silvan C, Rothschild-Mancinelli B, Balk M, Huttenhower C, Nocker A, Waishampayam P, Rothschild LJ (2017) Schrödinger’s microbes: tools for distinguishing the living from the dead in microbial ecosystems. Microbiome 5:1–23. https://doi.org/10.1186/s40168-017-0285-3
Fan J, Chen C, Brlansky RH, Gmitter FG, Li ZG (2010) Changes in carbohydrate metabolism in Citrus sinensis infected with ‘Candidatus Liberibacter asiaticus’. Plant Pathol 59:1037–1043. https://doi.org/10.1111/j.1365-3059.2010.02328.x
Gaire S, Albrecht U, Batuman O, Qureshi J, Zekri M, Alferez F (2022) Individual protective covers (IPCs) to prevent Asian citrus psyllid and Candidatus Liberibacter asiaticus from establishing in newly planted citrus trees. Crop Prot 152:105862. https://doi.org/10.1016/j.cropro.2021.105862
Giles F (2024) Florida citrus industry survey reflects industry optimism. Citrus Ind Magazine 105:20–21
Gottwald TR, da Graça d, Bassanezi B (2007) Citrus huanglongbing: The pathogen and its impact. Plant Health Prog 8. https://doi.org/10.1094/PHP-2007-0906-01-RV
Graham J, Gottwald T, Setamou M (2020) Status of Huanglongbing (HLB) outbreaks in Florida, California and Texas. Trop Plant Pathol 45:265–278. https://doi.org/10.1007/s40858-020-00335-y
Hall DG, Richardson ML, Ammar ED, Halbert SE (2013) Asian citrus psyllid, Diaphorina citri, vector of citrus huanglongbing disease. Entomol Exp Appl 146:207–223. https://doi.org/10.1111/eea.12025
Hamonts K, Trivedi P, Garg A, Janitz C, Grinyer J, Holford P, Botha FC, Anderson IC, Singh BK (2018) Field study reveals core plant microbiota and relative importance of their drivers. Environ Microbiol 20:124–140. https://doi.org/10.1111/1462-2920.14031
Hijaz F, Nehela Y, Batuman O, Killiny N (2021) Detection of oxytetracycline in citrus phloem and xylem saps using europium-based method. Antibiotics 10:1036. https://doi.org/10.3390/antibiotics10091036
Holochová P, Mašlaňová I, Sedláček I, Švec P, Králová S, Kovařovic V, Busse HJ, Staňková E, Barták M, Pantůček R (2020) Description of Massilia rubra sp. nov., Massilia aquatica sp. nov., Massilia mucilaginosa sp. nov., Massilia frigida sp. nov., and one Massilia genomospecies isolated from Antarctic streams, lakes and regoliths. Syst Appl Microbiol 43(126112). https://doi.org/10.1016/j.syapm.2020.126112
Hu J, Wang N (2016) Evaluation of the spatiotemporal dynamics of oxytetracycline and its control effect against citrus huanglongbing via trunk injection. Phytopathology 106:1495–1503. https://doi.org/10.1094/phyto-02-16-0114-r
Hu J, Jiang J, Wang N (2018) Control of citrus huanglongbing via trunk injection of plant defense activators and antibiotics. Phytopathology 108:186–195. https://doi.org/10.1094/phyto-05-17-0175-r
Kalam S, Basu A, Ahmad I, Sayyed RZ, El-Enshasy HA, Dailin DJ, Suriani NL (2020) Recent understanding of soil acidobacteria and their ecological significance: A critical review. Front Microbiol 11:580024. Recent understanding of soil acidobacteria and their ecological significance: A critical revie
Killiny N, Gonzalez-Blanco P, Santos-Ortega Y, Al-Rimawi F, Levy A, Hijaz F, Albrecht U, Batuman O (2019) Tracing penicillin movement in citrus plants using fluorescence-labeled penicillin. Antibiotics 8:262. https://doi.org/10.3390/antibiotics8040262
Killiny N, Hijaz F, Gonzalez-blanco P, Jones SE, Pierre MO, Vincent CI (2020) Effect of adjuvants on oxytetracycline uptake upon foliar application in citrus. Antibiotics 9:677. https://doi.org/10.3390/antibiotics9100677
Lareen A, Burton F, Schäfer P (2016) Plant root-microbe communication in shaping root microbiomes. PLant Mol Biol 90:575–587. https://doi.org/10.1007/s11103-015-0417-8
Leff JW, Del Tredici P, Friedman WE, Fierer N (2015) Spatial structuring of bacterial communities within individual Ginkgo biloba trees. Environ Microbiol 17:2352–2361. https://doi.org/10.1111/1462-2920.12695
Li J, Pang Z, Duan S, Lee D, Kolbasov VG, Wang N (2019) The in Planta Effective Concentration of Oxytetracycline against ‘Candidatus Liberibacter Asiaticus’ for suppression of Citrus Huanglongbing. Phytopathology 109:2046–2054. https://doi.org/10.1094/phyto-06-19-0198-r
Li L, Yang X, Tong B, Wang D, Tian X, Liu J, Chen J, Xiao X, Wang S (2023) Rhizobacterial compositions and their relationships with soil properties and medicinal bioactive ingredients in Cinnamomum migao. Front Microbiol 14:1078886. https://doi.org/10.3389/fmicb.2023.1078886
Lombardino J, Bijlani S, Singh NK, Wood JM, Barker R, Gilroy S, Wang CCC, Venkateswaran K (2022) Genomic characterization of potential plant growth-promoting features of Sphingomonas strains isolated from the international space station. Microbiol Spectr 10:e0199421. https://doi.org/10.1128/spectrum.01994-21
Maza-Márquez P, Castellano-Hinojosa A, González-Martínez A, Juárez-Jiménez B, González-López J, Rodelas B (2019) Abundance of total and metabolically active Candidatus Microthrix and fungal populations in three full-scale wastewater treatment plants. Chemosphere 232:26–34. https://doi.org/10.1016/j.chemosphere.2019.05.149
McVay J, Sun X, Jones D, Urbina H, Aldeek F, Cook JM, Jeyaprakash A, Hodges G, Smith T (2019) Limited persistence of residues and metabolites in fruit and juice following penicillin trunk infusion in citrus affected by Huanglongbing. Crop Prot 125:104753. https://doi.org/10.1016/j.cropro.2019.03.001
Morgan KT, Rouse RE, Ebel RC (2016) Foliar applications of essential nutrients on growth and yield of ‘valencia’ sweet orange infected with Huanglongbing. HortScience 51:1482–1493. https://doi.org/10.21273/HORTSCI11026-16
Nascimento FX, Tavares MJ, Glick BR, Rossi MJ (2018) Improvement of Cupriavidus taiwanensis nodulation and plant growth promoting abilities by the expression of an exogenous acc deaminase gene. Curr Microbiol 75:961–965. https://doi.org/10.1007/s00284-018-1474-4
Pal G, Saxena S, Kumar K, Verma A, Sahu PK, Pandey A, White JF, Verma SK (2022) Endophytic Burkholderia: multifunctional roles in plant growth promotion and stress tolerance. Microbiol Res 265:127201. https://doi.org/10.1016/j.micres.2022.127201
Qessaoui R, Bouharroud R, Furze JN, El Aalaoui M, Akroud H, Amarraque A, Vaerenbergh J, Van, Tahzima R, Mayad EH, Chebli B (2019) Applications of new rhizobacteria Pseudomonas isolates in agroecology via fundamental processes complementing plant growth. Sci Rep 9:1–10. https://doi.org/10.1038/s41598-019-49216-8
Rai AN, Singh AK, Syiem MB (2019) Plant growth-promoting abilities in Cyanobacteria. Cyanobacteria 459–476. https://doi.org/10.1016/B978-0-12-814667-5.00023-4
Raths R, Peta V, Bücking H (2021) Duganella callida sp. nov., a novel addition to the Duganella Genus, isolated from the soil of a cultivated maize field. Int J Syst Evol Microbiol 71:1–9. https://doi.org/10.1099/ijsem.0.004599
Riederer M (2004) Uptake and transport of xenobiotics. In: Hocl B, Elster EF (eds) Plant Toxicology. CRC, Boca Raton, FL, pp 131–150
Rodriguez-Sanchez C, Altendorf K, Smalla K, Lipski A (2008) Spraying of oxytetracycline and gentamicin onto field-grown coriander did not affect the abundance of resistant bacteria, reistance genes, and broad host range plasmids detected in tropical soil bacteria. Biol Fertil Soils 44:589–596. https://doi.org/10.1007/s00374-007-0242-6
Schepper V, De Swaef T, Bauweraerts I, Steppe K (2013) Phloem transport: a review of mechanisms and controls. J Exp Bot 64:4839–4850. https://doi.org/10.1093/jxb/ert302
Shin K, Ascunce MS, Narouei-Khandan HA, Sun X, Jones D, Kolawole OO, Goss EM, van Bruggen AHC (2016) Effects and side effects of penicillin injection in huanglongbing affected grapefruit trees. Crop Prot 90:106–116. https://doi.org/10.1016/j.cropro.2016.08.025
Sousa E, Naves P, Vieira M (2013) Prevention of pine wilt disease induced by Bursaphelenchus Xylophilus and Monochamus Galloprovincialis by trunk injection of emamectin benzoate. Phytoparasitica 41:143–148. https://doi.org/10.1007/s12600-012-0272-y
Stansly PA, Arevalo HA, Qureshi JA, Jones MM, Hendricks K, Roberts PD, Roka FM (2014) Vector control and foliar nutrition to maintain economic sustainability of bearing citrus in Florida groves affected by huanglongbing. Pest Manag Sci 70:415–426. https://doi.org/10.1002/ps.3577
Stoeck T, Bass D, Nebel M, Christen R, Jones MDM, Breiner H-W, Richards TA (2010) Multiple marker parallel tag environmental DNA sequencing reveals a highly complex eukaryotic community in marine anoxic water. Mol Ecol 19:21–31. https://doi.org/10.1111/j.1365-294X.2009.04480.x
Takahashi S, Tomita J, Nishioka K, Hisada T, Nishijima M (2014) Development of a prokaryotic universal primer for simultaneous analysis of bacteria and archaea using next-generation sequencing. PLoS ONE 9:e105592. https://doi.org/10.1371/journal.pone.0105592
Tardivo C, Qureshi J, Bowman KD, Albrecht U (2023) Relative influence of rootstock and scion on Asian citrus psyllid infestation and candidatus liberibacter asiaticus colonization. HortScience 58:395–403. https://doi.org/10.21273/HORTSCI17039-22
Trivedi P, Mattupalli C, Eversole K, Leach JE (2021) Enabling sustainable agriculture through understanding and enhancement of microbiomes. New Phytol 230:2129–2147. https://doi.org/10.1111/nph.17319
Verhaegen M, Bergot T, Liebana E, Stancanelli G, Streissl F, Mingeot-Leclercq MP, Mahillon J, Bragard C (2023) On the use of antibiotics to control plant pathogenic bacteria: a genetic and genomic perspective. Front Micro 14:1221478. https://doi.org/10.3389/fmicb.2023.1221478
Vincent CI, Hijaz F, Pierre M, Killiny N (2022) Systemic uptake of oxytetracycline and streptomycin in huanglongbing-affected citrus groves after foliar application and trunk injection. Antibiotics 11:1092. https://doi.org/10.3390/antibiotics11081092
Xu J, Zhang Y, Zhang P, Trivedi P, Riera N, Wang Y, Liu X, Fan G, Tang J, Coletta-Filho HD, Cubero J, Deng X, Ancona V, Lu Z, Zhong B, Rope MC, Capote N, Catara V, Pietersen G, Vernière C, Al-Sadi AM, Li L, Xu X, Wang J, Yang H, Jin T, Wang N (2018) The structure and function of the global citrus rhizosphere microbiome. Nat Commun 9:9:1–10. https://doi.org/10.1038/s41467-018-07343-2
Xu M, Jiang J, Feng Y, Li X, Ye L, Chen Y (2023) Resistance profiles of microbial communities in maize rhizospheres to the introduction of exogenous antibiotics to agricultural systems with a high arsenic geological background. Environ Sci Process Impacts 25:1224–1237. https://doi.org/10.1039/D3EM00149K
Yang Q, Zhang J, Zhu K, Zhang H (2009) Influence of oxytetracycline on the structure and activity of microbial community in wheat rhizosphere soil. J Environ Sci 21:954–959. https://doi.org/10.1016/S1001-0742(08)62367-0
Zhang M, Powell CA, Guo Y, Benyon L, Duan Y (2013) Characterization of the microbial community structure in Candidatus Liberibacter asiaticus-infected citrus plants treated with antibiotics in the field. BMC Microbiol 13:1–10. https://doi.org/10.1186/1471-2180-13-112
Zhang Z, He H, Han T, Tian X, Pang J, Lambers H (2023) Soil oxytetracycline alters the effects of phosphate fertilisation and Bacillus amyloliquefaciens on the bacterial community of Medicago sativa rhizosphere. Appl Soil Ecol 187:104861. https://doi.org/10.1016/j.apsoil.2023.104861
Acknowledgements
We thank the UF/IFAS SWFREC Plant Physiology and Soil Microbiology programs for their help with data and sample collection, and the grower collaborator for providing trees, tree care, and help with the harvest.
Funding
Funding for this project was provided by the Citrus Research and Development Foundation (22 − 001), USDA NIFA SCRI CDRE (2019-70016-29096), and USDA NIFA Hatch projects FLA-SWF-006160 and FLA-SWF-7000945.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Castellano-Hinojosa, A., González-López, J., Tardivo, C. et al. Trunk injection of oxytetracycline improves plant performance and alters the active bark and rhizosphere microbiomes in huanglongbing-affected citrus trees. Biol Fertil Soils 60, 563–576 (2024). https://doi.org/10.1007/s00374-024-01824-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00374-024-01824-x