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Heat Treatment Eliminates ‘Candidatus Liberibacter asiaticus’ from Infected Citrus Trees Under Controlled Conditions

First, second, sixth, seventh, and fourteenth authors: United States Department of Agriculture–Agriculture Research Service, United States Horticultural Research Laboratory, Fort Pierce, FL 34945; third, ninth, tenth, and twelfth authors: Division of Plant Industry, Florida Department of Agriculture and Consumer Services, Gainesville, FL 32614; fourth author: Institute of Food and Agricultural Science, Indian River Research and Education Center, University of Florida, Fort Pierce, FL 34945; fifth and eighth authors, Institute of Food and Agricultural Science, Tropical Research and Education Center, University of Florida, Homestead, FL 33031; and eleventh and thirteenth authors: Plant Pathology Department, University of Florida, Gainesville, FL 32614.

Huanglongbing (HLB) is one of the most destructive diseases of citrus worldwide. The three known causal agents of HLB are species of α-proteobacteria: ‘Candidatus Liberibacter asiaticus’, ‘Ca. L. africanus’, and ‘Ca. L. americanus’. Previous studies have found distinct variations in temperature sensitivity and tolerance among these species. Here, we describe the use of controlled heat treatments to cure HLB caused by ‘Ca. L. asiaticus’, the most prevalent and heat-tolerant species. Using temperature-controlled growth chambers, we evaluated the time duration and temperature required to suppress or eliminate the ‘Ca. L. asiaticus’ bacterium in citrus, using various temperature treatments for time periods ranging from 2 days to 4 months. Results of quantitative polymerase chain reaction (qPCR) after treatment illustrate significant decreases in the ‘Ca. L. asiaticus’ bacterial titer, combined with healthy vigorous growth by all surviving trees. Repeated qPCR testing confirmed that previously infected, heat-treated plants showed no detectable levels of ‘Ca. L. asiaticus’, while untreated control plants remained highly infected. Continuous thermal exposure to 40 to 42°C for a minimum of 48 h was sufficient to significantly reduce titer or eliminate ‘Ca. L. asiaticus’ bacteria entirely in HLB-affected citrus seedlings. This method may be useful for the control of ‘Ca. Liberibacter’-infected plants in nursery and greenhouse settings.