Publications

4592

‘Candidatus Liberibacter asiaticus’ (CLas) is a devastating pathogen of citrus associated with Huanglongbing (HLB, citrus greening disease). HLB is economically significant in Asia and has destroyed millions of citrus trees worldwide during the last century. Since 2007, when the first molecular evidence for the presence of CLas in North-West Frontier Province of Pakistan (Khyber Pakhtunkhwa) were presented, there have been a limited number of studies reporting the pathogen in different citrus growing districts of the Punjab Province, one of the major citrus producing areas of the country. In this study, a small-scale survey was conducted in citrus groves exhibiting HLB-like symptoms in different districts of Punjab Province. The aim was to obtain current information on the incidence of CLas in the area and complement the previous studies. Conventional and real time quantitative polymerase chain reaction (PCR and qPCR) assays were used for the detection of CLas in the 94 samples tested, followed by sequencing of the PCR amplicons. Overall CLas was detected in 43% (40/94) of the tested samples from Kinnow mandarin (n= 22), sweet orange (n= 12), grapefruit (n= 3) and Feutrell’s early mandarin (n= 3). In agreement with previous reports, this survey confirmed the presence of HLB in the Punjab Province of Pakistan and provided updated molecular evidence for the presence of CLas and disease incidence for four citrus cultivars from five citrus growing districts of the province.

ABSTRACT Ubiquitin‐proteasome is a conserved mechanism that regulates cellular responses and disease resistance in plants. However, the regulatory role of ubiquitin‐proteasome in the pathogenicity of “ Candidatus Liberibacter asiaticus” ( C Las), the causal agent of citrus huanglongbing (HLB), one of the most serious citrus diseases, remains poorly defined. In this study, we demonstrated that CLIBASIA_04250 (SDE4250) suppresses 26S proteasome activity in citrus, leading to a reduction in salicylic acid (SA) levels and downregulation of SA‐responsive genes. Further investigation revealed that SDE4250 specifically targets the citrus 26S proteasome non‐ATPase regulatory subunit 4 homologue (CsRPN10) and enhances its degradation via the 26S proteasome pathway. Moreover, overexpression of CsRPN10 inhibited C Las proliferation in citrus hairy roots, which was accompanied by a significant accumulation of SA content and upregulation of related defence genes. Collectively, our findings indicate that SDE4250 targets the citrus 26S proteasome to disrupt SA‐mediated citrus immunity.

Citrus huanglongbing (HLB), associated with the bacterium ‘Candidatus Liberibacter asiaticus’ and spread by the Asian citrus psyllid (Diaphorina citri; ACP), poses a significant threat to California’s citrus industry. First identified in Los Angeles in 2012, HLB has since spread through residential areas across Southern California. A risk-based survey (RBS) model has been developed to improve HLB surveillance and intervention. Within this framework, model components change as HLB dynamics shift, requiring regular updates to maintain data accuracy and model reliability. Disease spread is influenced by natural factors, such as ACP establishment and confirmed HLB locations, as well as human-mediated factors like global mobility (travel introduction from HLB-infected countries), transportation of citrus materials, nurseries, packinghouses, farmers’ markets, and proximity to private or otherwise inaccessible lands. Human-mediated risk factors account for approximately 26.3% (18.4 to 38.4%) of HLB incidence across different years, and natural causes predominantly explain the remaining 73.7% (61.6 to 81.7%). Notably, global mobility was crucial for early HLB detection in new areas, whereas ACP density strongly correlated with disease spread once established. A retrospective analysis from 2015 to 2022 evaluated the RBS model’s performance, showing a predictive power of 88 to 97%, which confirms its validity for developing targeted interventions and early detection strategies in California.

Citrus huanglongbing (HLB), caused by ‘Candidatus Liberibacter asiaticus’ (CLas), is a destructive disease threatening global citrus industry. Although citrus cultivars differ in HLB sensitivity, how infection alters endophytic bacterial communities in cultivars with contrasting susceptibility remains unclear. Here, we compared endophytic microbiome shifts in leaf and root tissues of HLB-susceptible Shatangju mandarin (Citrus reticulata cv. Shatangju) and HLB-tolerant Shatian pomelo (C. maxima cv. Shatian) at 10 months postgrafting with CLas-infected buds. Infected Shatangju mandarin showed severe symptoms and higher CLas levels in leaves than in roots, while infected Shatian pomelo remained asymptomatic with higher CLas in roots than in leaves. High-throughput 16S rRNA gene sequencing revealed that CLas infection restructured the endophytic microbial community in a tissue-specific manner. CLas-infected Shatian pomelo leaves displayed increased microbial diversity and network complexity, while CLas-infected Shatangju mandarin roots harbored more disease-suppressive taxa like Streptomyces. Across both cultivars, network complexity correlated inversely with CLas abundance. Key genera (e.g., Pseudomonas, Streptomyces, and Prevotella) correlated positively with CLas, suggesting roles in mitigating pathogen effects. Predicted functional profiling indicated activated pathways (amino acid metabolism, terpenoid/polyketide biosynthesis, and xenobiotic biodegradation) potentially supporting host defense in infected tissues. The HLB tolerance of Shatian pomelo appeared linked to tissue-specific bacterial restructuring, particularly recruitment of antibiotic-producing bacteria and enhanced metabolic resilience. These findings elucidated cultivar-specific microbial strategies against CLas, offering insights for microbiome-mediated HLB management.

Abstract Environmental genomics has led to the discovery of many new lineages of archaea, including “DPANN” (or Nanobdellati), comprising organisms with small genomes, reduced gene content, and potentially symbiotic or parasitic lifestyles. DPANN live in various environments, and several lineages have been identified that are adapted to extremely high-salt concentrations, including the Nanohaloarchaeota. Since it was long thought that the Haloarchaea (within “Euryarchaeota”) were the only high salt-adapted archaea, the origins of these genome-reduced halophiles have been debated. Here, we used phylogenetic, comparative genomic, and gene tree-species tree reconciliation approaches to resolve the evolution of halophily within DPANN, making use of recently published genomes that help to inform the phylogenetic placement and genome evolution of salt-adapted lineages. Phylogenetic analysis placed Nanohaloarchaeota sister to a previously uncharacterized lineage, which we here refer to as Terrarchaeota. Terrarchaeota appear to be predominantly anaerobic thermophiles that are not adapted to high-salt concentrations, indicating that adaptation to high salt evolved after their divergence from Nanohaloarchaeota. Furthermore, our analyses identified genomic hallmarks of salt adaptation in another recently discovered halophilic DPANN lineage within Aenigmatarchaeota, the Haloaenigmatarchaeaceae. We found that the Nanohaloarchaeota and Haloaenigmatarchaeaceae have distinct sets of proteins that enable life at high salt concentrations but share a common mechanism of evolutionary adaptation, in which niche-relevant genes were acquired horizontally from their halophilic hosts. This work provides the first detailed investigation into the enigmatic Terrarchaeota, and new insights into the convergent evolution of high salt adaptation within symbiotic clades of Archaea.

ABSTRACT Background Data on feline hemotropic mycoplasmas is limited in Iran. Aim. The aim of this study was to investigate the infection rate of cats with this pathogen in Mashhad,Iran and evaluate its assosiation with cliniopathological changes. Materias & Methods Blood samples from 100 cats presenting at veterinary clinics were analysed for Mycoplasma haemofelis (Mhf) and Candidatus Mycoplasma haemominutum (CMhm). Samples were screened for hemotropic mycoplasmas using microscopic examination and conventional polymerase chain reaction (PCR). Samples were also subjected to haematological and biochemical analysis. Results Microscopic examination detected hemotropic mycoplasmas in 65% of samples, whereas PCR identified a 23% infection rate, with 65.21% ( n  = 15) positive for CMhm and 34.78% ( n  = 8) for Mhf. The presence of Haemomycoplasma DNA was significantly associated with age, gender, breed and roaming status ( p  < 0.05). Infected cats exhibited significant alterations ( p  < 0.05) in haematocrit (Hct), red blood cell count (RBC), haemoglobin (HGB), white blood cell count (WBC), neutrophil counts, serum total protein (TP), globulin, phosphorus (Pi), aspartate aminotransferase (AST) levels and the albumin/globulin ratio (AGR). Conclusions Our study highlights a high prevalence of feline hemoplasmosis in northeastern Iran, with CMhm as the predominant species. Key risk factors included male gender, adult, outdoor access and domestic short hair (DSH) breed. Although infected cats showed consistent clinicopathologic changes, no differences were observed between Mhf and CMhm infections.