scholarly journals HPE1, an Effector from Zebra Chip Pathogen Interacts with Tomato Proteins and Perturbs Ubiquitinated Protein Accumulation

2021 ◽  
Vol 22 (16) ◽  
pp. 9003
Author(s):  
Chia-Cheng Kan ◽  
Azucena Mendoza-Herrera ◽  
Julien Levy ◽  
J. Joe Hull ◽  
Jeffery A. Fabrick ◽  
...  
Keyword(s):  
Direct Interaction ◽  
Ubiquitin Proteasome System ◽  
Hypothetical Protein ◽  
Protein Accumulation ◽  
In Planta ◽  
Zebra Chip ◽  
Citrus Greening Disease ◽  
Ubiquitinated Proteins ◽  
Candidatus Liberibacter ◽  
Effector Triggered Immunity

The gram-negative bacterial genus Liberibacter includes economically important pathogens, such as ‘Candidatus Liberibacter asiaticus’ that cause citrus greening disease (or Huanglongbing, HLB) and ‘Ca. Liberibacter solanacearum’ (Lso) that cause zebra chip disease in potato. Liberibacter pathogens are fastidious bacteria transmitted by psyllids. Pathogen manipulation of the host’ and vector’s immune system for successful colonization is hypothesized to be achieved by Sec translocon-dependent effectors (SDE). In previous work, we identified hypothetical protein effector 1 (HPE1), an SDE from Lso, that acts as a suppressor of the plant’s effector-triggered immunity (ETI)-like response. In this study, using a yeast two-hybrid system, we identify binding interactions between tomato RAD23 proteins and HPE1. We further show that HPE1 interacts with RAD23 in both nuclear and cytoplasmic compartments in planta. Immunoblot assays show that HPE1 is not ubiquitinated in the plant cell, but rather the expression of HPE1 induced the accumulation of other ubiquitinated proteins. A similar accumulation of ubiquitinated proteins is also observed in Lso infected tomato plants. Finally, earlier colonization and symptom development following Lso haplotype B infection are observed in HPE1 overexpressing plants compared to wild-type plants. Overall, our results suggest that HPE1 plays a role in virulence in Lso pathogenesis, possibly by perturbing the ubiquitin-proteasome system via direct interaction with the ubiquitin-like domain of RAD23 proteins.

scholarly journals Challenges for Managing Candidatus Liberibacter spp. (Huanglongbing Disease Pathogen): Current Control Measures and Future Directions

2018 ◽  
Vol 108 (4) ◽  
pp. 424-435 ◽  
Author(s):  
Ryan A. Blaustein ◽  
Graciela L. Lorca ◽  
Max Teplitski
Keyword(s):  
Broad Spectrum ◽  
Field Studies ◽  
Current Control ◽  
Control Measures ◽  
In Planta ◽  
Citrus Industry ◽  
Future Directions ◽  
Disease Symptoms ◽  
Citrus Greening Disease ◽  
Candidatus Liberibacter

Huanglongbing (HLB; “citrus greening” disease) has caused significant damages to the global citrus industry as it has become well established in leading citrus-producing regions and continues to spread worldwide. Insecticidal control has been a critical component of HLB disease management, as there is a direct relationship between vector control and Candidatus Liberibacter spp. (i.e., the HLB pathogen) titer in HLB-infected citrus trees. In recent years, there have been substantial efforts to develop practical strategies for specifically managing Ca. Liberibacter spp.; however, a literature review on the outcomes of such attempts is still lacking. This work summarizes the greenhouse and field studies that have documented the effects and implications of chemical-based treatments (i.e., applications of broad-spectrum antibiotics, small molecule compounds) and nonchemical measures (i.e., applications of plant-beneficial compounds, applications of inorganic fertilizers, biological control, thermotherapy) for phytopathogen control. The ongoing challenges associated with mitigating Ca. Liberibacter spp. populations at the field-scale, such as the seasonality of the phytopathogen and associated HLB disease symptoms, limitations for therapeutics to contact the phytopathogen in planta, adverse impacts of broad-spectrum treatments on plant-beneficial microbiota, and potential implications on public and ecosystem health, are also discussed.

scholarly journals HopA1 Effector from Pseudomonas syringae pv syringae Strain 61 Affects NMD Processes and Elicits Effector-Triggered Immunity

2021 ◽  
Vol 22 (14) ◽  
pp. 7440
Author(s):  
Shraddha K. Dahale ◽  
Daipayan Ghosh ◽  
Kishor D. Ingole ◽  
Anup Chugani ◽  
Sang Hee Kim ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Disease Susceptibility ◽  
Null Mutant ◽  
In Planta ◽  
Host Range Expansion ◽  
Unique System ◽  
Effector Triggered Immunity ◽  
Transcriptomic Changes ◽  
Immune Detection

Pseudomonas syringae-secreted HopA1 effectors are important determinants in host range expansion and increased pathogenicity. Their recent acquisitions via horizontal gene transfer in several non-pathogenic Pseudomonas strains worldwide have caused alarming increase in their virulence capabilities. In Arabidopsis thaliana, RESISTANCE TO PSEUDOMONAS SYRINGAE 6 (RPS6) gene confers effector-triggered immunity (ETI) against HopA1pss derived from P. syringae pv. syringae strain 61. Surprisingly, a closely related HopA1pst from the tomato pathovar evades immune detection. These responsive differences in planta between the two HopA1s represents a unique system to study pathogen adaptation skills and host-jumps. However, molecular understanding of HopA1′s contribution to overall virulence remain undeciphered. Here, we show that immune-suppressive functions of HopA1pst are more potent than HopA1pss. In the resistance-compromised ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1) null-mutant, transcriptomic changes associated with HopA1pss-elicited ETI are still induced and carry resemblance to PAMP-triggered immunity (PTI) signatures. Enrichment of HopA1pss interactome identifies proteins with regulatory roles in post-transcriptional and translational processes. With our demonstration here that both HopA1 suppress reporter-gene translations in vitro imply that the above effector-associations with plant target carry inhibitory consequences. Overall, with our results here we unravel possible virulence role(s) of HopA1 in suppressing PTI and provide newer insights into its detection in resistant plants.

scholarly journals Viability of ‘Candidatus Liberibacter asiaticus’ Prolonged by Addition of Citrus Juice to Culture Medium

2014 ◽  
Vol 104 (1) ◽  
pp. 15-26 ◽  
Author(s):  
Jennifer K. Parker ◽  
Sarah R. Wisotsky ◽  
Evan G. Johnson ◽  
Faraj M. Hijaz ◽  
Nabil Killiny ◽  
...  
Keyword(s):  
Grapefruit Juice ◽  
Culture Medium ◽  
Quantitative Polymerase Chain Reaction ◽  
Chemical Characterization ◽  
Candidatus Liberibacter Asiaticus ◽  
Citrus Greening Disease ◽  
Candidatus Liberibacter ◽  
The Media ◽  
Liberibacter Asiaticus

Huanglongbing, or citrus greening disease, is associated with infection by the phloem-limited bacterium ‘Candidatus Liberibacter asiaticus’. Infection with ‘Ca. L. asiaticus’ is incurable; therefore, knowledge regarding ‘Ca. L. asiaticus’ biology and pathogenesis is essential to develop a treatment. However, ‘Ca. L. asiaticus’ cannot currently be successfully cultured, limiting its study. To gain insight into the conditions conducive for growth of ‘Ca. L. asiaticus’ in vitro, ‘Ca. L. asiaticus’ inoculum obtained from seed of fruit from infected pomelo trees (Citrus maxima ‘Mato Buntan’) was added to different media, and cell viability was monitored for up to 2 months using quantitative polymerase chain reaction in conjunction with ethidium monoazide. Media tested included one-third King's B (K), K with 50% juice from the infected fruit, K with 50% commercially available grapefruit juice, and 100% commercially available grapefruit juice. Results show that juice-containing media dramatically prolong viability compared with K in experiments reproduced during 2 years using different juice sources. Furthermore, biofilm formed at the air–liquid interface of juice cultures contained ‘Ca. L. asiaticus’ cells, though next-generation sequencing indicated that other bacterial genera were predominant. Chemical characterization of the media was conducted to discuss possible factors sustaining ‘Ca. L. asiaticus’ viability in vitro, which will contribute to future development of a culture medium for ‘Ca. L. asiaticus’.

Differences in Zebra Chip Severity between ‘Candidatus Liberibacter Solanacearum’ Haplotypes in Texas

2018 ◽  
Vol 96 (1) ◽  
pp. 86-93 ◽  
Author(s):  
Kyle Harrison ◽  
Cecilia Tamborindeguy ◽  
Douglas C. Scheuring ◽  
Azucena Mendoza Herrera ◽  
Adrian Silva ◽  
...  
Keyword(s):  
Zebra Chip ◽  
Candidatus Liberibacter Solanacearum ◽  
Candidatus Liberibacter

scholarly journals A Yeast-Based Functional Assay to Study Plant N-Degron – N-Recognin Interactions

2022 ◽  
Vol 12 ◽  
Author(s):  
Aida Kozlic ◽  
Nikola Winter ◽  
Theresia Telser ◽  
Jakob Reimann ◽  
Katrin Rose ◽  
...  
Keyword(s):  
Ubiquitin Proteasome System ◽  
Functional Assay ◽  
The Novel ◽  
In Planta ◽  
Amino Terminal ◽  
Weak Binding ◽  
Ubiquitin Conjugating Enzyme ◽  
Ubiquitin Proteasome ◽  
The Stability

The N-degron pathway is a branch of the ubiquitin-proteasome system where amino-terminal residues serve as degradation signals. In a synthetic biology approach, we expressed ubiquitin ligase PRT6 and ubiquitin conjugating enzyme 2 (AtUBC2) from Arabidopsis thaliana in a Saccharomyces cerevisiae strain with mutation in its endogenous N-degron pathway. The two enzymes re-constitute part of the plant N-degron pathway and were probed by monitoring the stability of co-expressed GFP-linked plant proteins starting with Arginine N-degrons. The novel assay allows for straightforward analysis, whereas in vitro interaction assays often do not allow detection of the weak binding of N-degron recognizing ubiquitin ligases to their substrates, and in planta testing is usually complex and time-consuming.

scholarly journals Population Dynamics of Released Potato Psyllids and their Bacteriliferous Status in Relation to Zebra Chip Incidence in Caged Field Plots

Plant Disease ◽  
2016 ◽  
Vol 100 (8) ◽  
pp. 1762-1767 ◽  
Author(s):  
F. Workneh ◽  
L. Paetzold ◽  
A. Rashed ◽  
C. M. Rush
Keyword(s):  
Disease Incidence ◽  
Sticky Trap ◽  
Research Station ◽  
Zebra Chip ◽  
Sticky Traps ◽  
Monitoring Tools ◽  
Candidatus Liberibacter ◽  
Yellow Sticky Trap ◽  
Successive Generations ◽  
Field Plots

Potato psyllids vector ‘Candidatus Liberibacter solanacearum’ (Lso), the putative causal agent of potato zebra chip (ZC). Currently, sticky traps are the primary psyllid monitoring tools used by growers for making management decisions. However, the reliability of sticky traps in predicting psyllid numbers in potato fields has always been questioned. In 2013 and 2014, experiments were conducted in covered field plots at the Texas A&M AgriLife Research Station at Bushland to investigate the relationships among initial psyllid numbers, psyllids captured on sticky traps and their Lso status, and zebra chip incidence. Three densities of Lso-positive psyllids (5, 15, or 30/cage) were released under 2-week-old potato canopies with four replications in plot sizes of 7.6 to 9 m by 5 rows. Psyllids were released under the first plant in the center row and monitored weekly with a yellow sticky trap from the opposite end. Number of plants with zebra chip symptoms also was counted weekly beginning one month after infestation with psyllids. The total number of psyllids captured on sticky traps and disease incidence levels generally corresponded to the levels of psyllid density treatments (5 < 15 < 30), but the differences became more apparent toward the end of the experiments. Psyllid numbers in the different density treatments fluctuated more or less in synchrony over time, which appeared to reflect periodic emergence of new generations of psyllids. Initially, all captured psyllids tested positive for Lso. However, the proportions of psyllids testing positive declined dramatically after a few weeks, which suggested that the new generations of psyllids were devoid of Lso. Over all, less than 50% of captured psyllids tested positive for the pathogen. The decline in proportions of psyllids testing positive for Lso following successive generations has significant relevance to field situations and may partly explain why there are generally low percentages of Lso-positive psyllids under field conditions.

Probing the application of OmpA-derived peptides to disrupt the acquisition of ‘Candidatus Liberibacter asiaticus’ by Diaphorina citri

2021 ◽  
Author(s):  
Marcus Vinicius Merfa e Silva ◽  
Eduarda Regina Fischer ◽  
Mariana de Souza e Silva ◽  
Carolina Sardinha Francisco ◽  
Helvécio Coletta-Filho ◽  
...  
Keyword(s):  
Control Strategies ◽  
Insect Vector ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
In Planta ◽  
Host Interaction ◽  
Vector Population ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Bacterial Outer Membrane

Huanglongbing (HLB) is currently the most devastating disease of citrus worldwide. Both bacteria ‘Candidatus Liberibacter asiaticus’ (CLas) and ‘Ca. Liberibacter americanus’ (CLam) are associated with HLB in Brazil, but with a strong prevalence of CLas over CLam. Conventionally, HLB management focuses on controlling the insect vector population (Diaphorina citri; also known as Asian citrus psyllid – ACP) by spraying insecticides, an approach demonstrated to be mostly ineffective. Thus, development of novel more efficient HLB control strategies is required. The multifunctional bacterial outer membrane protein OmpA is involved in several molecular processes between bacteria and their hosts and has been suggested as a target for bacterial control. Curiously, OmpA is absent in CLam in comparison to CLas, suggesting a possible role on host-interaction. Therefore, in the current study, we have treated ACPs with different OmpA-derived peptides aiming to evaluate the acquisition of CLas by the insect vector. Treatment of psyllids with 5 µM of Pep1, Pep3, Pep5 and Pep6 in artificial diet significantly reduced the acquisition of CLas, while increasing the concentration of Pep5 and Pep6 to 50 µM abolished this process. In addition, in planta treatment with 50 µM of Pep6 also significantly decreased the acquisition of CLas and sweet orange plants stably absorbed and maintained this peptide for as long as three months post the final application. Together, our results demonstrate the promising use of OmpA-derived peptides as a novel biotechnological tool to control CLas.

scholarly journals The in Planta Effective Concentration of Oxytetracycline Against ‘Candidatus Liberibacter asiaticus’ for Suppression of Citrus Huanglongbing

2019 ◽  
Vol 109 (12) ◽  
pp. 2046-2054 ◽  
Author(s):  
Jinyun Li ◽  
Zhiqian Pang ◽  
Shuo Duan ◽  
Donghwan Lee ◽  
Vladimir G. Kolbasov ◽  
...  
Keyword(s):  
Field Experiments ◽  
Foliar Spray ◽  
Effective Control ◽  
Candidatus Liberibacter Asiaticus ◽  
In Planta ◽  
Fresh Tissue ◽  
Minimum Concentration ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Residue Levels

Huanglongbing (HLB) or greening currently is the most devastating citrus disease worldwide. The fastidious phloem-colonizing bacterium ‘Candidatus Liberibacter asiaticus’ (CLas) is the causal agent of citrus HLB in Florida. Bactericides containing the active ingredient oxytetracycline (OTC) have been used in foliar spray to control citrus HLB in Florida since 2016. However, the minimum concentration of OTC required to suppress CLas in planta remains unknown. We developed a new method for evaluating the effects of OTC treatment on CLas titers in infected plants and determined the relationship between OTC residue levels and control levels achieved for CLas using mathematical modeling in greenhouse and field experiments. In both greenhouse and field, OTC spray did not reduce the titers of CLas, and it produced undetectable or mild levels of OTC residue in leaves within 7 days post-application (DPA). In greenhouse, OTC injection at 0.05 g per tree decreased CLas titers to an undetectable level (cycle threshold value ≥ 36.0) from 7 to 30 DPA and produced a residue level of OTC at 0.68 to 0.73 µg/g of fresh tissue over this period. In the field, OTC injection at 0.50 g per tree resulted in the decline of CLas titers by 1.52 log reduction from 14 to 60 DPA, with residue levels of OTC at 0.27 to 0.33 µg/g of fresh tissue. In both trials, a first-order compart model of OTC residue dynamics in leaves of trunk-injected trees was specified for estimating the retention of effective concentrations. Furthermore, nonlinear modeling revealed significant positive correlations between OTC residue levels in leaves and the control levels for CLas achieved. The results suggested that the minimum concentrations of OTC required to suppress CLas populations in planta to below the detection limit are 0.68 and 0.86 µg/g and that the minimum concentrations of OTC required for initial inhibition of CLas growth in planta are ∼0.17 and ∼0.215 µg/g in leaf tissues under greenhouse and field conditions, respectively. This finding highlights that a minimum concentration of OTC should be guaranteed to be delivered to target CLas in infected plants for effective control of citrus HLB.

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