Leaf-disc grafting for the transmission of Candidatus Liberibacter asiaticus in citrus (Citrus sinensis; Rutaceae) seedlings1

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Abstract

Premise of the study:

The search for resistance/tolerance to the devastating citrus huanglongbing disease (syn. HLB or citrus greening) is generating an increasing number of new plants of diverse genetic makeup. As the increasing number of new plants require more space, resources, and time, the need for faster and more efficient HLB screening tests becomes crucial.

Methods and Results:

The leaf-disc grafting system described here consists in replacing a disc of leaf tissue with a similar disc from an infected plant. This can be performed in young seedlings not yet big enough to endure other types of grafting. Graft success and infection rates average approximately 80%.

Conclusions:

We describe the successful adaptation of leaf-disc grafting as a powerful screening tool for HLB. The system requires minimal plant material and can be performed in seedlings at a very young age with increased efficiency in terms of time, space, and resources.

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Most cited references 2

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Molecular and physiological mechanisms regulating tissue reunion in incised plant tissues.

Shinobu Satoh, Masashi Asahina (2015)

Interactions among the functionally specialized organs of higher plants ensure that the plant body develops and functions properly in response to changing environmental conditions. When an incision or grafting procedure interrupts the original organ or tissue connection, cell division is induced and tissue reunion occurs to restore physiological connections. Such activities have long been observed in grafting techniques, which are advantageous not only for agriculture and horticulture but also for basic research. To understand how this healing process is controlled and how this process is initiated and regulated at the molecular level, physiological and molecular analyses of tissue reunion have been performed using incised hypocotyls of cucumber (Cucumis sativus) and tomato (Solanum lycopersicum) and incised flowering stems of Arabidopsis thaliana. Our results suggest that leaf gibberellin and microelements from the roots are required for tissue reunion in the cortex of the cucumber and tomato incised hypocotyls. In addition, the wound-inducible hormones ethylene and jasmonic acid contribute to the regulation of the tissue reunion process in the upper and lower parts, respectively, of incised Arabidopsis stems. Ethylene and jasmonic acid modulate the expression of ANAC071 and RAP2.6L, respectively, and auxin signaling via ARF6/8 is essential for the expression of these transcription factors. In this report, we discuss recent findings regarding molecular and physiological mechanisms of the graft union and the tissue reunion process in wounded tissues of plants.

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Transmission and Propagation of 'Candidatus Liberibacter asiaticus' by Grafting with Individual Citrus Leaves.

Mark Hilf, Reid Lewis (2016)

Huanglongbing (HLB) is a chronic, progressive decline disease in citrus associated with a systemic infection by the bacterium 'Candidatus Liberibacter asiaticus'. Transmission of the bacterium in the field is by the Asian citrus psyllid, Diaphorina citri Kuwayama. Experimental propagation of 'Ca. L. asiaticus' is done primarily by grafting pieces of bud wood from an infected plant. To produce a small-scale model system for investigation of pathogen biology, we investigated grafting single leaves from infected citrus plants as sources of inoculum for propagation of the bacterium. In total, 162 plants ranging in age from 3 to 18 months were grafted. Grafting with intact asymptomatic and HLB-symptomatic leaves resulted in 61 of 78 (78%) and 35 of 41 (85%) of the plants infected with 'Ca. L. asiaticus', respectively. Inoculum consisting of the leaf petiole only or only an inoculum tissue remnant under the bark of the receptor tree resulted in 6 of 12 (50%) and 7 of 31 (23%) infected trees, respectively. Real-time polymerase chain reaction (qPCR) assays verified the infection in plants, a majority of which developed the foliar blotchy mottle symptom considered diagnostic for HLB, while some plants also displayed the stunted, chlorotic shoots for which the disease is named. The qPCR data together with the symptoms displayed demonstrated that individual leaves from infected trees can serve as effective inoculum sources for transmission and propagation of 'Ca. L. asiaticus' via grafting.

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Author and article information

Journal

Journal ID (nlm-ta): Appl Plant Sci

Journal ID (iso-abbrev): Appl Plant Sci

Journal ID (publisher-id): apps

Title: Applications in Plant Sciences

Publisher: Botanical Society of America

ISSN (Electronic): 2168-0450

Publication date Collection: January 2017

Publication date (Electronic): 5 January 2017

Volume: 5

Issue: 1

Electronic Location Identifier: apps.1600085

Affiliations

[2 ]Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred, Florida 33850 USA

Author notes

[1]

This work was supported in part by the Citrus Research Development Foundation (no. 15-008).

[3 ]Author for correspondence: eetxeber@ 123456ufl.edu

Article

Publisher ID: apps1600085

DOI: 10.3732/apps.1600085

PMC ID: 5231911

SO-VID: 9d638a85-fbc1-40f9-bd8a-403a4d518204

License:

This is an open access article distributed under the terms of the Creative Commons Attribution License ( CC-BY-NC-SA 4.0), which permits unrestricted noncommercial use and redistribution provided that the original author and source are credited and the new work is distributed under the same license as the original.

History

Date received : 18 July 2016

Date accepted : 6 December 2016

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