Two miniature inverted-repeat transposable elements (MITEs), MCLas-A and MCLas-B, were recently identified from ‘Candidatus Liberibacter asiaticus’ known to be associated with citrus Huanglongbing (HLB, yellow shoot disease). MCLas-A was suggested as an active MITE because of its mobility. The immediate upstream gene of the two MITEs was predicted to be a putative transposase. The goal of this study is to analyze the sequence variation in the upstream putative transposase of MITEs and explore the possible correlation between sequence variation of transposase gene and MITE activity. PCR and sequence analysis showed that 12 sequence types were found in six major amplicon types from 43 representative ‘Ca. L. asiaticus’ isolates from China, the United States and Brazil. Out of the 12 sequence types, three (T4, T5-2, T6) were reported for the first time. Recombination events were found in the two unique sequence types (T5-2 and T6) which were detected in all Brazilian isolates. Notably, no sequence variation or recombination events were detected in the upstream putative transposase gene of MCLas-A, suggesting the conservation of the transposase gene might be closely related with the MITE activity. Phylogenetic analysis demonstrated two well supported clades including five subclades were identified, clearly reflecting the geographical origins of isolates, especially that of Ruili isolates, São Paulo isolates and a few Florida isolates.

Citrus Huanglongbing (HLB, yellow shoot disease) was first observed in the coastal Chaoshan Plain of Guangdong Province, China, in the late 19th century based on descriptions of yellow shoot symptoms. “Candidatus Liberibacter asiaticus” has been considered as a putative pathogen associated with HLB since 1994. Information about the curent prevalence of this bacterium is important for HLB control in Guangdong and also provides useful reference for HLB study elsewhere. In 2007, we collected HLB symptomatic citrus samples from 16 cultivars in 12 prefecture cities, mostly in the north and west regions of Guangdong, where major citrus fruits are currently produced. Among the 359 samples collected, 241 (67.1%) were positive for “Ca. L. asiaticus”, distributed in 15 out of the 16 cultivars from all 12 cities, indicating the widespread prevalence of “Ca. L. asiaticus” in Guangdong Province. The detection rates varied from 16.7 to 100% depending on location and cultivar. Lower detection rates were found in newer citrus cultivation cities among the previously less popular but now promoted cultivars. In reviewing the citrus management and pest control practice, we believe that infected nursery stocks play a key role in the current spread of “Ca. L. asiaticus”.