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**Huanglongbing (Citrus Greening): What ARS Is Doing** Huanglongbing (HLB), also known as Citrus Greening disease because fruit tends to turn green after ripening, undoubtedly poses the most serious threat that the Florida citrus industry has ever faced. The bacteria that cause HLB—three species of Liberibacter—probably originated in China in the early 1900s. In countries where the disease is endemic, citrus trees begin to decline within 5 to 8 years after planting and rarely bear usable fruit. First detected in Florida in 2005; by 2008, it had been identified in most of the citrus growing counties in the state. Despite everyone's best efforts, HLB now literally threatens the survival of Florida citrus and is a potential threat to the entire U.S. citrus industry. ARS scientists are actively engaged in research with university and industry partners on the critical components of this disease such as etiology and epidemiology, the insect vector, and the plant host's response to HLB infection. In February 2008, members of the citrus industry requested additional input from ARS scientists, especially from researchers outside of the citrus field, to draw on the most diverse expertise possible to elicit new ideas about research needs, gaps, priorities, and opportunities. In response, ARS convened a workshop in April 2008 at the U.S. Horticultural Research Laboratory in Fort Pierce, Florida, especially inviting ARS scientists who are leaders in their scientific fields but who are not ordinarily engaged in citrus research. These ARS scientists brought their collective expertise in animal and plant entomology, animal and plant pathology, genetics, horticulture, and microbiology to the workshop. The workshop was coordinated with the National Academy of Sciences (NAS), which had been asked by the Florida Department of Citrus to develop a Request for Proposals to manage grant funding for research leading to HLB management. That meeting was followed in May 2008 by a workshop in Beltsville, Maryland, of key ARS citrus researchers to discuss potential new collaborations and research directions to address the needs identified at the April 2008 workshop.
 **Recent News** New Traps Cut Off Citrus Greening Pests from Hiding Places July 13, 2020Trained Dogs Are the Most Efficient Way to Hunt Citrus Industry's Biggest Threat February 3, 2020Research Reveals a New Direction for Halting the Citrus Greening Epidemic March 26, 2018
 On September 17-18, 2008, a group of ARS scientists, industry and university representatives from citrus producing states met with the federal regulatory agencies USDA-APHIS, FDA, and EPA to prepare for the advent of transgenic citrus to be used to combat HLB and canker to seek guidance on bringing these products to market.. This meeting was an excellent opportunity for each party to learn about what will be needed as progress is made toward this goal. Other potential new chemicals for insect control and management of the pathogen in citrus were also discussed. A team of scientists are preparing documents that summarize published data on citrus biology, pollination, gene flow and juice composition parameters; information that will be needed by the regulatory agencies as transgenic citrus products are developed. **Current ARS Research** ARS has initiated a significant research program, even before the 2008 workshops. ARS scientists have established partnerships with industry and university colleagues, and these collaborations continue to grow and diversity as new knowledge is developed. The ultimate goal is to effectively manage major threats to the citrus industry in the U.S., in particular, citrus greening, or HLB, and canker. A broad approach is being taken to target the main components of the disease triangle; the host, the pathogen and associated organisms, and the insect vector. Insect management, repellants and control are critical to disease control. Manipulating citrus groves by varying nutrition and high density planting may reduce the impact of the diseases. As scientists learn more about the bacterium associated with HLB, methods for directly impacting its growth in the citrus host will be devised. Conventional breeding and genetically modified citrus varieties are being explored. All of these components of disease intervention are being investigated by ARS scientists and colleagues, and will be utilized for to benefit the national citrus industry.
 Third Annual Citrus Health Research Forum International Research Conference on Huanglongbing USDA Public Citrus Genome Database First Annual Citrus Health Research Forum: Agenda and Presentations Outcome Summary Reports PostersJune 16-18, 2010
 **Accomplishments** Because of the fastidious nature of HLB bacteria (*Candidatus*Liberibacter asiaticus), a high-throughput detection technology for HLB is critical for HLB research and disease management. Polyclonal antibodies against HLB pathogen have been developed by researchers at Fort Pierce, Florida, and a series of primers and probes were also developed for better detection of HLB using PCR or in situ hybridization. These technologies have been transferred to university and state regulatory research laboratories for HLB detection and other research purposes. ARS scientists at Frederick, Maryland, determined the effect of imidacloprid insecticide on transmission of HLB. Following incorporation of the insecticide into sweet orange seedlings, infectious psyllids were aspirated on the seedlings and monitored for survival and disease transmission. Although most psyllids died within the first 24 hours, strong symptoms of HLB infection were evident by 8 weeks in most plants, indicating the insects could transmit the causal HLB bacterium before dying. ARS scientists at Beltsville, Maryland developed and validated the first real-time and quantitative PCR assay for the greening pathogen. This assay was used by the USDA Animal and Plant Health Inspection Service (APHIS) and the Florida Department of Agriculture to confirm the presence of HLB in Florida. Producers will benefit from early detection and genetic improvement for resistance will be accelerated. ARS scientists at Ft. Pierce and colleagues recently published the full genomic sequence of HLB bacterium, 'Candidatus Liberibacter asiaticus'. This is a major scientific advance, and will enable scientists to identify genes responsible for the disease, enhance capabilities to culture this organism and determine its true role as the causal agent.
 **For more information, contact:** MaryLou PolekCitrus Research Boardmarylou@citrusresearch.org559-738-0246 Gail C. WislerNational Program LeaderHorticulture and SugarGail.Wisler@ars.usda.gov(301) 504-4562.
 Scientists at Ft. Pierce have evidence that suggests that guava, interplanted with citrus, prevents the spread of greening. Scientists in Japan and Viet Nam discovered that HLB did not occur in citrus groves when intercropped with guava, while control plots had over 30 percent infection. The mechanism may be volatile compounds produced by guava which disrupt the ability of the psyllid vector of HLB to locate its citrus host. Studies are underway in field plots to validate these preliminary findings and to identify the potential volatiles. ARS Research Projects on Huanglongbing (Citrus Greening) Exotic Pathogens of CitrusLEAD SCIENTIST: John S. Hartung, Beltsville, Maryland IPM Technologies for Subtropical Insect PestsLEAD SCIENTIST: David Hall, Fort Pierce, Florida Identification, Characterization, and Biology of Foreign and Emerging Insect-Transmitted Plant PathogensLEAD SCIENTIST: Vernon D. Damsteegt, Ft. Detrick, Maryland Identification, Characterization, and Detection of Foreign and Newly Emerging Domestic BacteriaLEAD SCIENTIST: Norm Schaad, Ft. Detrick, Maryland Epidemiology and Management of Xylella Fastidiosa (XF) and other Exotic and Invasive Diseases and Insect PestsLEAD SCIENTIST: Drake Stenger, Parlier, California
 **For more information about ARS HLB programs, contact:** Gail C. WislerNational Program LeaderHorticulture and SugarGail.Wisler@ars.usda.gov(301) 504-4562. Deborah FravelActing National Program LeaderPlant Healthdeborah.fravel@ars.usda.gov(301) 504-5450
 Management of Citrus and Date Genetic Resources and InformationLEAD SCIENTIST: Richard Lee, Riverside, California Genetic Improvement of CitrusLEAD SCIENTIST: Kim Bowman, Fort Pierce, Florida Domestic, Exotic, and Emerging Diseases of Citrus, Vegetables, and Ornamentals (DEED)LEAD SCIENTIST: Timothy Gottwald, Fort Pierce, Florida ARS Research Publications on Citrus Greening/Huanglongbing (HLB) Publications from the USDA Horticultural Research Laboratory on the Asian Citrus Psyllid and Huanglongbing Disease Publication Abstract:Effect of Early Detection Greening on Juice Flavor and Chemistry Publication Abstract:NPDN Citrus Greening Diagnostic and Detection Efforts
 Asian Citrus Psyllid and Citrus Greening - A Closer Look at the VectorCitrus and Vegetable Magazine Huanglongbing, Citrus Greening: Preparing for the FutureSubtropics Newsletter Twelve Polymorphic Microsatellite Loci from the Asian Citrus Psyllid, Diaphorina Citri (Kuwayama), the Vector for Citrus Greening Disease (Huanglongbing)Molecular Ecology Notes Sensitivity Of PCR-based Methods for Detection of Asiatic Strain of the Huanglongbing Bacterium in Symptomatic and Asymptomatic Citrus TissuesProceedings of the Huanglongbing-greening International Workshop, July 14-21, 2006, Ribeiro Preto, Brazil Publication Abstract: Differences in Secondary Metabolites in Leaves from Trees Affected With the Greening (Hlb) Disease Citrus Huanglongbing: The Pathogen and Its ImpactPlant Health Progress Citrus Canker and Citrus Huanglongbing, Two Exotic Bacterial Diseases Threatening the Citrus Industries of the Western HemisphereOutlooks on Pest Management Progress Report to the Citrus Industry: Florida Citrus Plant Protection Committee (FCPPC)Citrus Industry Detection And Identification of Candidatus Liberobacter Species Associated with Citrus Huanglongbing by Multiplex Real-time PCRJournal of Microbiological Methods Gene Expression in the Asian Citrus Psyllid: Vector of Citrus Greening (Hemiptera: Psyllidae)Joint Proceedings of the 89th Annual meeting and 6th International Caribbean Conference of the Florida Entomologtical Society Incidence Of the Asian Citrus Psyllid, Diaphorina Citri (Hemiptera: Psyllidae) and its Parasitoid Tamarixia Radiata (Hymenoptera: Eulophidae) in Florida CitrusThe 91st Annual Meeting of the Florida Entomological Society Ribosomal Proteins of the Asian Citrus Psyllid, Diaphornia CitriNational Center for Biotechnology Information (NCBI) Reovirus-like Sequences Isolated from Adult Asian Citrus Psyllid, (Hemiptera: Psyllidae: Diaphorina Citri)Florida Entomologist Comparison Of Visual Assessment to Polymerase Chain Reaction Assay Testing to Estimate the Incidence of the Huanglongbing Pathogen in Commercial Plantings in FloridaProceedings of Florida State Horticultural Society
 More ARS information about HLB: ARS Workshop SummaryApril 2008 ARS Projects Funded by theFlorida Citrus Production ResearchAdvisory CouncilMay 2008
 Gene Expression in Citrus Sinensis (L.) Osbeck after Infection with the Bacterial Pathogen Candidatus Liberibacter Asiaticus Causing Huanglongbing in FloridaPlant Science Evaluation Of DNA Amplification Methods for Improved Detection of Candidatus Liberibacter Species Associated with Citrus HuanglongbingPlant Disease Publication AbstractGenome Sequence of Candidatus Liberibacter Asiaticus, the Casual Agent of Citrus Huanglongbing (Greening) Effects Of a Particle Film on Biology and Behavior of Diaphorina Citri (Hemiptera: Psyllidae) and its Infestations in CitrusJournal of Economic Entomology Discovery of a Viral Pathogen in the Asian Citrus Pysllid72nd Annual Meeting of the Florida Academy of Sciences in conjunction with the 85th Annual Meeting of the Georgia Academy of Sciences, March 14-15, Jacksonville, Florida Daily Timing of Mating and Age at Reproductive Maturity in Diaphorina Citri (Hemiptera: Psyllidae)Florida Entomologist Optimized Quantification of Unculturable Candidatus Liberibacter Species Causing Citrus Huanglongbing in Host Plants by Real-Time PCRPlant Disease
 Additional information about HLB: Citrus GreeningAnimal and Plant Health Inspection Service SaveOurCitrus.orgSave Our Citrus campaign (APHIS). Huanglongbing/Citrus Greening DiseaseFlorida Department of Agriculture and Consumer Services
 
 Publication Abstract:Genome Sequence of Candidatus Liberibacter Asiaticus, the Casual Agent of Citrus Huanglongbing (Greening) Publication Proceedings:Concepts in Huanglongbing Epidemiology Multiplex Taqman-based PCR for Sensitive and Accurate Quantification of Citrus HLB and CVC Pathogens; Candidatus Liberibacter and Xylella FastidiosaProceedings of the Huanglongbing-greening International Workshop Optimized Quantification of Unculturable Candidatus Liberibacter Species Causing Citrus Huanglongbing in Host Plants by Real-Time PCRPlant Disease First Report of Dodder Transmission of Huanglongbing from Naturally Infected Murraya paniculata to CitrusPlant Disease Incidence of Huanglongbing in Psyllids Under Natural Conditions in FloridaPhytopathology Preliminary Spatial Pattern Analysis of Huanglongbing in Sao Paulo, BrazilInternational Organization of Citrus Virologists Proceedings Use of Psyllids in Early Identification of Huanglongbing Disease of CitrusPhytopathology Huanglongbing Epidemiology: an International PerspectiveProceedings of Huanglongbing-greening International Workshop, July 14-21, 2006, Ribeiro Preto, Brazil Huanglongbing: The Dragon Arrives in the USAHuanglongbing-greening International Workshop, July 14-21,2006, Ribeiro Preto, Brazil
 Epidemiology of Huanglongbing in Sao PauloHuanglongbing-greening International Workshop, July 14-21, 2006, Ribeiro Preto, Brazil Huanglongbing Epidemiology: Tracking the Dragon Through Time and SpaceSecond International Citrus Canker and Huanglongbing Workshop, Orlando, FL, November 7-11, 2005 Spatial Distribution of Huanglongbing Symptomatic Trees in Citrus Groves in Sao Paulo, BrazilSecond International Citrus Canker and Huanglongbing Workshop, Orlando, FL, November 7-11, 2005 Detection Of Candidatus Liberibacter Asiaticus from Citrus Huanglongbing Samples in China By Nested Conventional and Real-time PCRPhytopathology Biology, History and World Status of Diaphorina CitriNorth American Plant Protection Organization Workshop, Taller Internacional Sobre Huanglongbing y el Psilido Asiatico de los Citricos, del 7 at 9 de Mayo de 2008, Hermosillo, Sonora
 Progress Towards the Development of an Effective Risk Analysis Process for the Florida Citrus Nursery Industry to Mitigate the Impact of Citrus Canker and HuanglongbingProceedings from the International Citrus Canker and Huanglongbing Workshop, Orlando, FL, Nov. 2005 Intercropping Of Citrus and Guava Trees for Management of HuanglongbingFlorida Entomological Society Annual Meeting, July 15-18, 2007, Sarasota, Florida Psyllids as a Tool in Evaluating the Efficiency of Different Management Practices for Control of Citrus Huanglongbing DiseasePhytopathology Novel Molecular Technique for Rapid Cloning of Unknown Sequences From Unculturable Huanglongbing (HLB) Associated Bacterium, Candidatus LiberibacterAmerican Phytopathology Society Meeting Murraya paniculata (Orange Jasmine), a Host and Possible Inoculum Reservoir for Candidatus Liberibacter Asiaticus, Causal Agent of HuanglongbingPhytopathology Large-scale Surveys for Multiple Pest Species; the Search for Citrus Canker and Huanglongbing in FloridaAmerican Phytopathological Society Annual Meeting Comparative Genomics and Phylogenetic Analyses of Newly Cloned Genomic Regions From the Citrus Huanglongbing (HLB)-associated Bacterium Candidatus LiberibacterAmerican Phytopathology Society


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