Publications

4560

The symptoms of huanglongbing (HLB), a disease caused by the bacterium Candidatus Liberibacter asiaticus (CLas), are visible on the canopy of citrus plants. They include mottling of leaves followed by dropping and lopsided fruits with premature dropping. Loss in phloem functionality and degradation of the root system are also HLB symptoms with a severe impact on plant growth and production. Some Citrus relatives, such as Poncirus trifoliata and its hybrids, have shown more tolerance to HLB disease and low titers of CLas compared to Citrus species, but little is known about the effects of CLas on their root system. In this study, we investigated the effects of CLas-infected ‘Valencia’ scion on the citrandarin IAC3222 (a hybrid between P. trifoliata and Sunki mandarin) used as rootstock as well as interstock between ‘Valencia’ and Swingle citrumelo rootstock. At 13 months post-inoculation, the cycle threshold values (CT) for CLas in the infected scion samples indicated a high CLas titer (from 15.9 to 22.7) regardless of the rootstock variety or interstock used. However, no CLas-positive samples were detected in the roots of IAC3222 (CT ranging from 37.9 to 40.0), in contrast to all Swingle roots (CT ranging from 27.9 to 31.3). Both root volume and mass were reduced in IAC3222 compared to uninfected ‘Valencia’ scion, suggesting that scion infection damages roots, regardless of whether they are contaminated or not by CLas. The damage to the root system of IAC3222 was significantly less severe than that of the Swingle rootstock. Multivariate hierarchical analysis considering all evaluated parameters clustered the CLas-infected plants grafted on IAC3222 together with the non-inoculated plants. We concluded that the IAC3222 rootstock was less affected by the CLas-infected scion compared to the Swingle rootstock and is a promising rootstock to minimize the HLB effect on plant development.

ABSTRACT‘CandidatusLiberibacter asiaticus’ (Las) is a gram-negative bacterial pathogen associated with citrus huanglongbing (HLB) or greening disease. Las is transmitted by the Asian citrus psyllid (ACP) where it colonizes the phloem tissue, resulting in substantial economic losses to citrus industry worldwide. Despite extensive efforts, effective management strategies against HLB remain elusive, necessitating a deeper understanding of the pathogen’ s biology. Las undergoes cell-to-cell movement through phloem flow and colonizes different tissues in which Las may have varying interactions with the host. Here, we investigate the transcriptomic landscape of Las in citrus seed coat vasculatures, enabling a complete gene expression profiling of Las genome and revealing unique transcriptomic patterns compared to previous studies using midrib tissues. Comparative transcriptomics between seed coat, midrib and ACP identified specific responses and metabolic states of Las in different host tissue. Two Las virulence factors that exhibit higher expression in seed coat can suppress callose deposition. Therefore, they may contribute to unclogging sieve plate pores during Las colonization in seed coat vasculature. Furthermore, analysis of regulatory elements uncovers a potential role of LuxR-type transcription factors in regulating Liberibacter effector gene expression during plant colonization. Together, this work provides novel insights into the pathogenesis of the devastating citrus HLB.FundingThis work is supported by USDA National Institute of Food and Agriculture award No. 2020-70029-33197 to W.M and A.L.

AbstractSpotted fever group (SFG) rickettsia, the causative agent of SFG rickettsiosis, is predominantly carried by ticks, whereas Orientia tsutusgamushi, the causative agent of scrub typhus, is primarily transmitted by chigger mites in Japan. In this study, we attempted to isolate intracellular eubacteria from Leptotrombidium scutellare, a major vector of O. tsutsugamushi; moreover, we isolated an SFG rickettsia using a mosquito‐derived cell line. Draft genome sequences of this unique isolate, by applying criteria for species delimitation, classified this isolate as a novel strain, proposed as “Rickettsia kedanie.” Further genetic analysis identified conserved virulence factors, and the isolate successfully propagated in mammalian cells, suggesting its ability to cause diseases in humans. The presence of SFG rickettsia in unfed larvae implies potential dual‐pathogen carriage and reflects a symbiotic relationship similar to that between the mites and O. tsutsugamushi, indicating possibility of its transovarial transmission from female adults. Furthermore, conserved genomic similarity of the novel isolate to known SFG rickettsia suggests potential multiple hosts, including chiggers and ticks. In the natural environment, ticks, chigger mites, and wild animals may carry new isolates, complicating the infection cycle and increasing the transmission risks to humans. This discovery challenges the conventional association of SFG rickettsia with ticks, emphasizing its implications for research and disease control. However, this study was confined to a particular species of chigger mites and geographic area, underscoring the necessity for additional studies to comprehend the ecological dynamics, host interactions, and health implications linked to this newly identified SFG rickettsia.

Abstract Citrus huanglongbing (HLB) has been causing enormous damage to the global citrus industry. As the main causal agent, ‘ Candidatus Liberibacter asiaticus’ ( C Las) delivers a set of effectors to modulate host responses, while the modes of action adopted remain largely unclear. Here, we demonstrated that CLIBASIA_00185 ( C Las0185) could attenuate reactive oxygen species (ROS)‐mediated cell death in Nicotiana benthamiana . Transgenic expression of C Las0185 in Citrus sinensis ‘Wanjincheng’ enhanced plant susceptibility to C Las. We found that methionine sulphoxide reductase B1 (CsMsrB1) was targeted by the effector, and its abundance was elevated in CLas0185 ‐transgenic citrus plants. Their interaction promoted C Las proliferation. We then determined that CsMsrB1 sustained redox state and enzymatic activity of ascorbate peroxidase 1 (CsAPX1) under oxidative stress. The latter reduced H 2 O 2 accumulation and was associated with host susceptibility to C Las infection. Consistently, citrus plants expressing C Las0185 and CsMsrB1 conferred enhanced APX activity and decreased H 2 O 2 content. Taken together, these findings revealed how C Las0185 benefits C Las colonization by targeting CsMsrB1, which facilitated the antioxidant activity and depressed ROS during pathogen infection.

Recurrent epiphytotics of X-disease, caused by ‘Candidatus Phytoplasma pruni,’ have inflicted significant losses on commercial cherry and peach production across North America in the last century. During this period, there have been multiple studies reporting different disease phenotypes and, more recently, identifying different strains through sequencing core genes, but the symptoms have not, to date, been linked with genotype. Therefore, in this study we collected and assessed differing disease phenotypes from multiple U.S. states and conducted multilocus sequence analysis on these strains. We identified a total of five lineages associated with the induction of X-disease on commercial Prunus species and two lineages that were associated with wild P. virginiana. Despite a century of interstate plant movement, there were regional trends in terms of lineages present, and lineage-specific symptoms were observed on P. avium, P. cerasus, and P. virginiana, but not on P. persica. Cumulatively, these data have allowed us to define “true” X-disease–inducing strains of concern to the stone fruit industry across North America, as well as potential sources of infection that exist in the extraorchard environment.