A stable antimicrobial peptide with dual functions of treating and preventing citrus Huanglongbing

Abstract

Significance Citrus Huanglongbing (HLB) is the most destructive citrus disease worldwide and has caused billions of dollars in annual production losses, threatening the entire citrus industry. Despite extensive research efforts, there are still no effective management tools to treat HLB-positive trees or to prevent new infections. Current HLB management strategies include chemical application of insecticides and traditional heat-sensitive antibiotics, which pose threats to humans, animal health, and the environment, and likely generate drug resistant insects and microbes. Here, we identified a novel class of stable antimicrobial peptides (SAMPs) from Australian finger lime and other HLB-tolerant citrus close relatives, which has dual functions of inhibiting CLas growth in HLB-positive trees and activating host immunity to prevent new infections. Citrus Huanglongbing (HLB), caused by a vector-transmitted phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas), is the most devastating citrus disease worldwide. Currently, there are no effective strategies to prevent infection or to cure HLB-positive trees. Here, using comparative analysis between HLB-sensitive citrus cultivars and HLB-tolerant citrus hybrids and relatives, we identified a novel class of stable antimicrobial peptides (SAMPs). The SAMP from Microcitrus australiasica can rapidly kill Liberibacter crescens (Lcr), a culturable Liberibacter strain, and inhibit infections of CLas and CL. solanacearum in plants. In controlled greenhouse trials, SAMP not only effectively reduced CLas titer and disease symptoms in HLB-positive trees but also induced innate immunity to prevent and inhibit infections. Importantly, unlike antibiotics, SAMP is heat stable, making it better suited for field applications. Spray-applied SAMP was taken up by citrus leaves, stayed stable inside the plants for at least a week, and moved systemically through the vascular system where CLas is located. We further demonstrate that SAMP is most effective on α-proteobacteria and causes rapid cytosol leakage and cell lysis. The α-helix-2 domain of SAMP is sufficient to kill Lcr. Future field trials will help determine the efficacy of SAMP in controlling HLB and the ideal mode of application.