Field Epidemiology of Huanglongbing: High Psyllid Density, Disease Severity, and an Alternative Host in Northern Thailand

Abstract

This study investigated the epidemiology and incidence of citrus Huanglongbing (HLB) disease in citron (Citrus medica L.) under varying field conditions. This research specifically aimed to quantify disease severity, assess populations of the primary vector, the Asian citrus psyllid (Diaphorina citri), and identify potential alternative host plants sustaining the vector and the pathogen. Field surveys were executed across three sites characterised by distinct elevations and management practices. Site-level soil nutrient profiles exhibited moderate acidity (pH 4.67–5.74) and significant differences in organic matter and nitrogen. These findings suggest that localised deficiencies in calcium and boron may exacerbate disease severity, contributing to the varied epidemiological patterns observed across sites. Analysis revealed that both HLB disease severity and D. citri population density were significantly influenced by altitude, field condition, and orchard management. The low-elevation site (860 m above sea level (ASL)), characterised by poor maintenance, exhibited the highest mean psyllid populations (averaging 13 individuals per sticky trap per day) and the most severe disease symptoms. Conversely, the high-elevation site (1674 m ASL) displayed significantly lower infection rates and healthier tree conditions. Symptomatic citron trees across all sites consistently exhibited characteristic HLB foliar and fruit symptoms (blotchy mottle and lopsided fruits). Quantitative Polymerase Chain Reaction (qPCR) successfully detected the causal agent, Candidatus Liberibacter asiaticus (CLas), in symptomatic citron samples from all locations, with the highest relative fold-change (2−ΔΔCt = 4628.24). Crucially, multiple developmental stages of D. citri were observed infesting the common weed Bidens pilosa. Furthermore, qPCR confirmed the presence of CLas DNA within the B. pilosa tissue itself (2−ΔΔCt = 210.84). This finding constitutes the first field-based evidence that B. pilosa can serve as a novel alternative host that supports both the D. citri vector and the CLas pathogen. These results establish citron as a highly susceptible host and identify B. pilosa as a new, critical epidemiological link in the HLB transmission cycle, thereby underscoring the necessity for integrated, landscape-level disease management strategies.