Publications

4693

‘ Candidatus Liberibacter asiaticus’ ( C Las), the causal agent of citrus huanglongbing, is transmitted by the Asian citrus psyllid Diaphorina citri . While C Las-positive ( C Las+) females exhibit increased fecundity and metabolic demands, their neuroendocrine regulation mechanisms remain unclear. We propose C Las manipulates dopamine (DA) signaling to enhance psyllid fecundity and C Las proliferation. Metabolomics revealed elevated DA in C Las+ females. Silencing DA synthesis genes and receptor DcDop2 via RNAi reduced lipid reserves, fecundity, and ovarian C Las titers. Through combined in vivo and in vitro experiments, we demonstrated that the microRNA miR-31a suppresses DcDop2 expression by binding to its 3’ untranslated region. Overexpression of miR-31a resulted in decreased DcDop2 expression and C Las titers in the ovaries, eliciting phenotypic defects akin to DcDop2 knockdown. Furthermore, DcDop2 knockdown and miR-31a overexpression reduced juvenile hormone (JH) levels and adipokinetic hormone (AKH) signaling in fat bodies and ovaries. Consequently, C Las regulates the DA- DcDop2 signaling axis to improve D. citri lipid metabolism and fecundity, while simultaneously promoting its replication. These findings reveal a coevolution between C Las proliferation and ovarian development in the insect host. This discovery enhances our understanding of the molecular interplay between plant pathogens and vector insects and offers novel targets and strategies for HLB field management.

‘ Candidatus Liberibacter asiaticus’ ( C Las), the causal agent of citrus huanglongbing, is transmitted by the Asian citrus psyllid Diaphorina citri . While C Las-positive ( C Las+) females exhibit increased fecundity and metabolic demands, their neuroendocrine regulation mechanisms remain unclear. We propose C Las manipulates dopamine (DA) signaling to enhance psyllid fecundity and C Las proliferation. Metabolomics revealed elevated DA in C Las+ females. Silencing DA synthesis genes and receptor DcDop2 via RNAi reduced lipid reserves, fecundity, and ovarian C Las titers. Through combined in vivo and in vitro experiments, we demonstrated that the microRNA miR-31a suppresses DcDop2 expression by binding to its 3’ untranslated region. Overexpression of miR-31a resulted in decreased DcDop2 expression and C Las titers in the ovaries, eliciting phenotypic defects akin to DcDop2 knockdown. Furthermore, DcDop2 knockdown and miR-31a overexpression reduced juvenile hormone (JH) levels and adipokinetic hormone (AKH) signaling in fat bodies and ovaries. Consequently, C Las regulates the DA- DcDop2 signaling axis to improve D. citri lipid metabolism and fecundity, while simultaneously promoting its replication. These findings reveal a coevolution between C Las proliferation and ovarian development in the insect host. This discovery enhances our understanding of the molecular interplay between plant pathogens and vector insects and offers novel targets and strategies for HLB field management.

Huanglongbing, a destructive citrus disease of global importance that is also present in Saudi Arabia, is associated with Candidatus Liberibacter asiaticus (CLas) and remains a major threat to citrus production. Although previous studies have documented sequence variation and prophage polymorphism in CLas, broader comparisons of prophage-associated gene content remain limited. In particular, comparative orthogroup analysis of prophage gene-content conservation across geographically structured CLas populations has rarely been explored. In this study, we analyzed 42 CLas prophage genomes from Saudi Arabia and other geographic regions using a comparative orthogroup framework. OrthoFinder assigned 99.1% of predicted proteins (1825 of 1841) to 64 orthogroups, with only 16 genes remaining unassigned. A small number of rare orthogroups restricted to only a few genomes were identified, and no orthogroup was detected in all genomes. Presence–absence analyses supported a predominantly conserved prophage gene repertoire together with a small accessory component, while also indicating that apparent absences should be interpreted in light of mixed assembly status and prophage-region completeness. Saudi Arabian genomes were distributed within the broader global framework and exhibited generally similar gene-content profiles rather than a deeply separated lineage. Functional interpretation of representative orthogroups identified conserved prophage-associated genes related to replication, helicase activity, and phage packaging, whereas variable orthogroups were primarily associated with hypothetical or accessory prophage-related functions. Overall, these results are consistent with a model in which CLas prophage diversification is associated more with sequence-level variation and localized structural differences than with extensive gain or loss of prophage genes. These findings further refine current understanding of CLas genome evolution and highlight conserved prophage-associated targets that may support molecular diagnostics and epidemiological surveillance.

Chokecherry (Prunus virginiana L.) is a native shrub that is widely found across North America and is a host of ‘Candidatus Phytoplasma pruni’ associated with X-disease of stone fruits. This pathogen is of major concern to U.S. cherry and peach growers, with the most recent and ongoing outbreak in the Pacific Northwest causing major economic harm. To investigate the role of chokecherry as a potential reservoir and source of orchard-associated strains, we surveyed wild chokecherry trees across 12 U.S. states, and compared the strains present to those in commercially cultivated Prunus species. Approximately 47% of chokecherry sampled tested positive for a ‘Ca. P. pruni’-related strain and phylogenetic analysis revealed high strain diversity, including six distinct clades and four newly identified lineages. In contrast, ‘Ca. P. pruni’ strains isolated from cultivated cherry and peach trees exhibited lower diversity and were infected by known disease outbreak-associated strains such as NWX. These data suggest limited transmission between wild and cultivated hosts in the western U.S., with patterns indicating spillover from orchards into nearby chokecherry populations rather than the reverse. However, in states east of the Rocky Mountains, chokecherry likely functions as a reservoir for orchard infections.

Abstract A comprehensive polyphasic taxonomic strategy was applied to the systematic characterization of strain DNPG3 T , which was isolated from the river Ganges, Hooghly, West Bengal, India. The Gram-positive, halotolerant, heavy-metal-tolerant strain exhibited the ability to degrade p -nitrophenol (PNP). Cellular fatty acid analysis revealed that the predominant components were anteiso-C 15:0 (24.61%), C 11:0 (21.06%), iso-C 16:0 (11.89%), C 16:0 (11.58%), and anteiso-C 17:0 (11.24%). Notably, the presence of C 11:0 , C 10:0 2-OH as major fatty acids differentiate strain DNPG3 T from its closely related members of the genus Brachybacterium . The predominant respiratory quinone was identified as menaquinone-7 (MK-7). Analysis of 16S rRNA gene sequence indicated that B. zhongshanense strain JB T was the closest relative of DNPG3 T , sharing 97.08% sequence similarity. Genome-based ANI value calculated using the EzBioCloud server revealed that B. zhongshanense JCM 15471 T was the closest genomic relative (85.49%). These values were further substantiated by digital DNA–DNA hybridization (dDDH) estimates calculated using the GGDC server. Taxonomic assignment using the GTDB database further indicated that strain DNPG3 T constitutes a previously unrecognized species within the genus Brachybacterium . Genome analysis of strain DNPG3 T identified eleven genomic islands, along with a rich repertoire of 194 carbohydrate-active enzyme (CAZyme) families, comprising 95 glycoside hydrolases and 53 glycosyltransferases. In addition, five biosynthetic gene clusters were detected. Collectively, these genomic features indicate the involvement of horizontal gene transfer events and highlighted the pronounced metabolic versatility of the strain, underscoring its potential for industrial enzyme production and secondary metabolite biosynthesis. Pan-genome analysis further indicates that the Brachybacterium pan-genome is open, reflecting substantial genetic diversity and ongoing gene acquisition within the genus. Comprehensive biochemical, physiological, chemotaxonomic, and phylogenetic analyses supported the assignment of strain DNPG3 T to the genus Brachybacterium while clearly distinguishing it from all currently described species within the genus. Accordingly, strain DNPG3 T was proposed to represent a novel species, for which the name Brachybacterium netajii sp. nov. is suggested. The type strain was DNPG3 T (= MTCC13125 T ).