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Biology subjects

Lovelace, A. H.

Publications and source records attributed to Lovelace, A. H..

4 recordsLinked to original sources

A secreted citrus protease cleaves an outer membrane protein of the Huanglongbing pathogen

Plants secrete a variety of proteases as a defense response during infection by microbial pathogens. However, the relationship between their catalytic activities and antimicrobial functions remains largely unknown. Particularly, few biologically relevant substrates of these proteases have been identified. Huanglongbing (HLB) has been a major threat to the citrus industry worldwide. The HLB-associated bacterium, Candidatus Liberibacter asiaticus (Las), was previously shown to deploy an inhibitor of papain-like cysteine proteases (PLCPs) to promote disease in citrus. In this study, we identified an outer membrane protein (OMP) of Las, LasOMP1, as a substrate of the citrus PLCP CsRD21a. LasOMP1 is one of the most highly expressed genes in Las. CsRD21a cleaves LasOMP1 and produces cleaved peptide products, which could be detected in vitro and in HLB-diseased citrus plants. We found that CsRD21a targets the N-terminal portion of LasOMP1, potentially at an extracellular loop region. Importantly, transgenic sweet orange overexpressing CsRD21a showed reduced Las titers and delayed disease symptoms, highlighting that engineering this protease is a promising strategy to enhance HLB resistance in citrus. Together, our work reveals a pathogen-derived substrate of plant PLCPs and highlights bacterial OMPs as direct targets of plant defense. Significance StatementHuanglongbing (HLB) is the most devastating disease of citrus with no resistance having been identified in commercial cultivars. Previous work implicated papain-like cysteine proteases (PLCPs) as an important hub of defense in citrus; however, their precise role in HLB tolerance remained unclear. Here, we identify and characterize an outer membrane protein (OMP) from the HLB-associated bacterium as a substrate of the citrus PLCP RD21a. We demonstrate a specific cleavage of the bacterial OMP by citrus RD21a, which may impair pathogen growth and/or activate plant immunity. Importantly, overexpression of RD21a enhances HLB tolerance in sweet oranges. This work identifies OMPs as substrates of plant PLCPs and provides insights into protease defense functions.

plant biology↗

Genome-wide association mapping and predictive modeling of wet bean mass in a diverse cacao collection

Improving cacao yield, a key objective in post-domestication crop improvement, remains a primary goal for breeders, but progress is often hindered by the confounding effects of population structure. To overcome this, we analyzed 346 diverse cacao accessions using an ML-based association mapping framework (with and without population structure adjustment) and a phenotype-only ML prediction of yield. By correcting for population structure, our Bootstrap Forest-based GWAS revealed association signals that showed consistent enrichment for ribosome and protein-synthesis functions, and a recurrent subset of SNPs with high importance appeared across multiple yield components, including pod index and seed number. In parallel, a Neural Network model was utilized to identify cotyledon mass and length as the most powerful predictors for total wet bean mass (R{superscript 2} = 0.715 by repeated five-fold cross-validation), suggesting a practical, low-cost screening proxy for breeding). Collectively, this study delivers a robust genetic framework and a novel predictive tool to accelerate the development of high-yielding cacao varieties through the early identification of elite clones.

genomics↗

Transcriptomic profiling of Candidatus Liberibacter asiaticus in different citrus tissues reveals novel insights into Huanglongbing pathogenesis

Candidatus Liberibacter 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.

microbiology↗

RpoS Contributes to Successful Opportunistic Colonization by Human Enteric Pathogens during Plant Disease

With an increase in foodborne illnesses associated with the consumption of fresh produce, it is important to understand the interactions between human bacterial enteric pathogens and plants. It was previously established that diseased plants can create a permissive environment for opportunistic endophytic colonization of enteric pathogens. However, the factors that contribute to the colonization of enteric pathogens during plant disease are largely unknown. Here, we show that both strain and plant host factors contribute to significantly increased populations of enteric pathogens when co-inoculated with the plant pathogen, P. syringae pv. tomato. The two Salmonella enterica strains DM10000 and 14028S, differ in their ability to metabolize host-derived apoplastic carbohydrates dependent on the sigma factor RpoS. The rpoS gene is an important strain factor for endophytic colonization by S. enterica during plant disease. Our results suggest that rpoS plays a crucial role during in planta colonization, balancing nutrient metabolism and stress responses. ImportanceFoodborne illnesses caused by the bacterial human enteric pathogens, E. coli O157:H7 and S. enterica, often results in vomiting and diarrhea. If left untreated, this illness can cause dehydration and sometimes death of a patient. Both E. coli O157:H7 and S. enterica have caused repeated fresh produce-associated epidemics. Crop disease could promote the ability of plants to act as reservoirs for produce-borne outbreaks. Plant pathogens dampen plant immunity, which allows for a more permissive environment for human enteric pathogens to grow. These internalized enteric pathogen populations are especially dangerous since they cannot be removed by washing alone. Therefore, the need to understand the factors that contribute to the opportunistic colonization of human enteric pathogens during plant disease is apparent. Our research has identified host and strain factors that contribute to opportunistic colonization of diseased plants, which will inform the development of future management strategies to mitigate future outbreaks.

pathology↗