Identification of a chromosomal deletion mutation and the dynamics of two major populations of Candidatus Liberibacter asiaticus in its hosts

Candidatus Liberibacter asiaticus (Las) is the prominent species of Liberibacter associated with huanglongbing, a devastating disease of citrus worldwide. In this study, we report the identification of a ~8.3 kb DNA region of the Las genome containing eight putative open reading frames (ORFs) flanked by two inverted repeats, which was not present in the Las str. psy62 genome. Comparisons with other genome sequences established this region as a unique genetic element associated with genome plasticity/instability. Primers specific for both the presence (Las wild-type) and absence (Las mutant) of this region were designed to study the population dynamics and host adaptation of the two strains. Las populations with and/or without the wild-type strain were detected and differentiated in >2,300 samples that included psyllids, periwinkle, and several species of citrus. In psyllids, although a mixed population of the wild-type and mutant was observed in most samples (88%), the wild-type Las was detected alone at a rate of 11%. In contrast, none of the infected citrus plants were positive for the wild-type alone, which harbored either the mutant strain alone (8%) or a mixed population of the mutant and wild-type (92%). Furthermore, the dynamics of these two major Las populations varied with different citrus hosts while an in-depth study on grapefruit that did not rapidly succumb to disease revealed that the population of mutant alone increased with time, indicating that the absence of this genetic element is associated with the fitness of Las in planta under the selection pressure of its host.