2011 Annual Report 1a.Objectives (from AD-416) 1) Identify genetic diversity of HLB pathogen(s) in China and the United States. 2) Identify molecular and biological mechanisms of citrus HLB and control of the disease 1b.Approach (from AD-416) Identify and characterize the disease-free citrus plant (s) from historically and heavily HLB-affected orchard (over 10-15 year old citrus plants). Isolate and identify the microbial consortium from the rhizosphere and endophytes from the identified disease-free plants by culture the microbes and sequencing the microbes using molecular markers. Characterize the role of Can. L asiaticus in the HLB disease development using functional genomics. Characterize the host gene expression and regulation of the citrus plants those are resistant or tolerant to HLB by using microarray and real-time PCR. Characterize the biological and molecular mechanisms of HLB bacterial transmission by the Asian Citrus psyllid using comparative and functional genomics. Explore and apply the biocontrol potentials of the psyllids by using biocontrol microorganism and parasitic or predator insects. Screening chemicals against HLB and application of the selected chemical in the fields and monitoring the dynamics of the disease in the field. 3.Progress Report This project is related to inhouse objective 1: Characterize ecology, biology, epidemiology, genetics and host interactions of domestic, exotic, newly emergent and re-emerging pathogens. The project is currently at the third year. Genetic diversity of the Huanglongbing (HLB) pathogens in China and the United State Samples from various HLB-infected citrus varieties were collected both from China and the United States, and the DNAs were isolated and subjected to genetic diversity studies based on the genetic markers identified from the genome sequence of the HLB bacterium, ‘Candidatus Liberibacter asiaticus’(Las). Sequence analysis of the hyvI and hyvII genes of these isolates indicated their genetic diversity between these two origins, which suggested the Las strain in Florida is different from those in China. Furthermore, Interclade assignments of the putative HyvI/II proteins from Florida isolates with other global isolates in phylogenetic trees imply multiple Las populations in the world, and a multi-source introduction of the Las bacterium into Florida. Progress was monitored via through direct involvement in lab and field activities, research meetings, telephone calls and email communications.