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Molecular and cellular analysis of orange plants infected with Huanglongbing (citrus greening disease)

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Abstract

Huanglongbing (HLB, also commonly known as citrus greening disease) is caused by the bacterium Candidatus Liberibacter asiaticus (CLas) in the US. It is a serious plant disease causing tremendous economic damages and posing an existential threat to the citrus industry worldwide. CLas proliferates inside the phloem, clogs the transportation system, stunts plant growth and renders trees barren. Currently there is no cure or effective treatments for this disease. In this study, qPCR and scanning electron microscopy (SEM) were used to examine orange plants with or without HLB symptoms. We found that symptomatic HLB plants showed high levels of CLas DNA and its prophage in leaves and had heavily clogged phloem tissues. In contrast, the asymptomatic, healthy looking plants had detectable, low levels of CLas and prophage DNA by qPCR but the SEM imaging revealed phloem clear of clogging deposits. Transcription of several CLas phage genes was detected in both healthy and diseased plants, however, higher transcription levels were observed in the healthy plants than in the diseased plants. As part of our efforts to explore the possibility of using a plant-based antibacterial concoction named Agent G to mitigate HLB, NMR and GC/MS were used to assess its organic chemical composition and migration through the phloem tissue. It was found that the major compounds of Agent G were allyl polysulfides and cinnamaldehyde. The phloemic migrate rate in planta was estimated between 1.5 and 3.25 cm/h. Our studies suggest that a combination of qPCR test and SEM imaging may prove to be more informative for gauging growth status of citrus plants and for early HLB detection. We also showcase a feasible approach of treating HLB by delivering antibacterial reagents directly into the phloem tissue where most CLas reside.

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Acknowledgements

The scanning electron microscopy work was funded by and carried out in A.D. Henderson University High School, Florida Atlantic University Owls Imaging Lab with technical assistance from Andrea Hernandez.

Funding

This work was supported in part by a fund from Penta5, USA. NP and MA were awarded a Florida Atlantic University undergraduate research grant.

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Authors and Affiliations

  1. Department of Biological Sciences, Florida Atlantic University, Boca Raton, FL, 33431, USA

    Xing-Hai Zhang, Nicholas Pizzo, Mohamed Abutineh & Xiao-Lu Jin

  2. College of Education, Florida Atlantic University, Boca Raton, FL, 33431, USA

    Nicholas Pizzo, Mohamed Abutineh & Tricia L. Meredith

  3. Department of Chemistry and Biochemistry, Florida Atlantic University, Boca Raton, FL, 33431, USA

    Sarah Naylon & Lyndon West

  4. Penta5, USA, Sarasota, FL, 34234, USA

    John M. Harlin

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  1. Xing-Hai Zhang
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  4. Xiao-Lu Jin
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Contributions

XHZ and JMH conceived and designed the experiments. XLJ and XHZ performed DNA and RNA analyses. XHZ, NP, MA and TLM performed scanning electron microscopy. LW, SN and XHZ performed composition and phloem migration analyses. XHZ, LW and JMH analyzed the data and wrote the manuscript. All commented on the manuscript.

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Correspondence to Xing-Hai Zhang or John M. Harlin.

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Zhang, XH., Pizzo, N., Abutineh, M. et al. Molecular and cellular analysis of orange plants infected with Huanglongbing (citrus greening disease). Plant Growth Regul 92, 333–343 (2020). https://doi.org/10.1007/s10725-020-00642-z

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