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.

Population diversity of “Candidatus Liberibacter asiaticus” and Diaphorina citri in Sichuan: A case study for Huanglongbing monitoring and interception

Plant Disease ◽  
2021 ◽  
Author(s):  
Xuejin Cui ◽  
Kehong Liu ◽  
Jie Huang ◽  
Shimin Fu ◽  
Qingdong Chen ◽  
...  
Keyword(s):  
Rural Areas ◽  
Urban Areas ◽  
Control Strategies ◽  
Population Diversity ◽  
Mitochondrial Genomes ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Multiple Sources ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

Citrus Huanglongbing (HLB) is present in 10 provinces in China and is associated with “Candidatus Liberibacter asiaticus” (CLas), which is transmitted by the Asian citrus psyllid (Diaphorina citri, ACP). To date, HLB and ACP have expanded to Yibin city of Sichuan Province, posing an imminent threat to the citrus belt of upper and middle reach of Yangtze River, an important late maturing citrus-producing area in China. To understand the epidemiological route of CLas and ACP in newly invaded regions of Sichuan and thereby better establish an HLB-interception zone ranging from Leibo to Yibin, we evaluated the molecular variability of 19 CLas draft genomes from citrus or dodder (Cuscuta campestris). They include three type-specific prophage loci, three variable numbers of tandem repeat (VNTR) loci, a miniature inverted-repeat transposable element (MITE) types, and population diversity of 44 ACP mitochondrial genomes. The results indicated that CLas isolates in the newly invaded area (Pingshan) were more diverse than those in the HLB endemic areas (Leibo and Ningnan). Phylogenetic analysis based on mitochondrial genomes demonstrated that ACPs in Leibo, Pingshan and Xuzhou (rural areas) represent a new group (MG4), distinguished by the three unique SNPs in cox1, nad4 and cytb. However, the ACPs sampled from the urban areas of Cuiping and Xuzhou belonged to the southeastern China group (MG2-1). Altogether, our study revealed multiple sources of ACP and CLas in the HLB-interception zone and proposed their transmission route. This study contributes to the formulation of precise HLB prevention and control strategies in the HLB-interception zone in Sichuan and could be useful for HLB management efforts in other regions.

Prevalent transmission of 'Candidatus Liberibacter asiaticus’ over ‘Ca. Liberibacter americanus’ in a long-term controlled environment

2021 ◽  
Author(s):  
Maria Cândida de Godoy Gasparoto ◽  
Isabela Vescove Primiano ◽  
Renato Bassanezi ◽  
Silvia Afonseca Lourenço ◽  
Luiz Montesino ◽  
...  
Keyword(s):  
Sweet Orange ◽  
Insect Vector ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Candidatus Liberibacter ◽  
Citrus Huanglongbing ◽  
Liberibacter Asiaticus ◽  
Citrus Orchards ◽  
Test Plants

In Brazil, citrus huanglongbing (HLB) is associated with ‘Candidatus Liberibacter americanus’ (CLam) and ‘Ca. Liberibacter asiaticus’ (CLas). However, there are few studies about HLB epidemiology when both Liberibacter spp. and its insect vector, the Asian citrus psyllid (ACP, Diaphorina citri), are present. The objective of this work was to compare the transmission of HLB by ACP when both CLam and CLas are present as primary inoculum. Two experiments were performed under screenhouse conditions from April 2008 to January 2012 (experiment 1) and from February 2011 to December 2015 (experiment 2). The experiments were carried out with sweet orange plants infected with CLam or CLas as inoculum source surrounded by sweet orange healthy plants. One hundred Liberibacter-free adult psyllids were monthly confined to the source of inoculum plants for 7 days with subsequent free movement inside the screenhouse. Fortnightly, nymphs and adults of psyllids were monitored. Psyllid and leaf samples were collected periodically for Liberibacter detection by PCR or qPCR. CLas was detected more frequently than CLam in both psyllid and leaf samples. No mixed infections were detected in the psyllids. A clear prevalence of CLas over CLam was observed in both experiments. The final HLB incidences were 16.7 and 14.5% of Liberibacter-positive test plants, and CLas was detected in 92.3 and 93.1% of these infected plants. Mixed infection was observed only in 3.8% of infected test plants in experiment 1. These results endorse the shift in the prevalence of CLam to CLas observed in citrus orchards of São Paulo, Brazil.

scholarly journals Genome Sequence Resource of ‘Candidatus Liberibacter asiaticus’ from Diaphorina citri Kuwayama (Hemiptera: Liviidae) in Colombia

Plant Disease ◽  
2021 ◽  
Vol 105 (1) ◽  
pp. 193-195
Author(s):  
Yanjing Wang ◽  
Takumasa Kondo ◽  
Yurong He ◽  
Zhuohuai Zhou ◽  
Jinming Lu
Keyword(s):  
South America ◽  
Genome Sequence ◽  
Insect Vector ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Introduction Pathway ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

‘Candidatus Liberibacter asiaticus’ (CLas) is an unculturable phloem-restricted α-proteobacterium associated with huanglongbing (HLB). Here, we provide the genome sequence of CLas strain CoFLP1 from its insect vector Diaphorina citri (Hemiptera: Liviidae) collected in the department of La Guajira, Colombia. The CoFLP1 strain is composed of 1,231,639 bp with G+C 36.5% content. This study reports the first CLas genome sequence from Colombia, which will add to CLas genome resources and help to elucidate our understanding of the introduction pathway of HLB in South America.

Distribution and Variation of Bacterial Endosymbiont and “Candidatus Liberibacter asiaticus” Titer in the Huanglongbing Insect Vector, Diaphorina citri Kuwayama

2018 ◽  
Vol 78 (1) ◽  
pp. 206-222 ◽  
Author(s):  
Saeed Hosseinzadeh ◽  
Masoud Shams-Bakhsh ◽  
Marina Mann ◽  
Somayeh Fattah-Hosseini ◽  
Abdoolnabi Bagheri ◽  
...  
Keyword(s):  
Insect Vector ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Bacterial Endosymbiont ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

scholarly journals Genome Sequence Resource of ‘Candidatus Liberibacter asiaticus’ from Diaphorina citri Kuwayama (Hemiptera: Liviidae) from La Réunion

Plant Disease ◽  
2020 ◽  
Author(s):  
Jinming Lu ◽  
Hélène Delatte ◽  
Bernard Reynaud ◽  
George Andrew Charles Beattie ◽  
Paul Holford ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Whole Genome Sequence ◽  
Insect Vector ◽  
Whole Genome ◽  
Diaphorina Citri ◽  
La Réunion ◽  
Candidatus Liberibacter Asiaticus ◽  
Introduction Pathway ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

‘Candidatus Liberibacter asiaticus’ (CLas) is an insect-transmitted, phloem-restricted α-proteobacterium associated with huanglongbing (HLB). Here, we provide the whole genome sequence of CLas strain, ReuSP1, from its insect vector Diaphorina citri (Hemiptera: Liviidae) collected in La Réunion. The genome is composed of 1,230,064 bp and has a 36.5% G+C content. This study reports the first CLas genome sequence from La Réunion, which will add to CLas genome resources and help elucidate our understanding of the introduction pathway into La Réunion.

scholarly journals Uptake, Translocation, and Stability of Oxytetracycline and Streptomycin in Citrus Plants

Antibiotics ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 196 ◽  
Author(s):  
Fuad Al-Rimawi ◽  
Faraj Hijaz ◽  
Yasser Nehela ◽  
Ozgur Batuman ◽  
Nabil Killiny
Keyword(s):  
Enzyme Linked Immunosorbent Assay ◽  
Asian Citrus Psyllid ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
In Planta ◽  
Citrus Industry ◽  
Candidatus Liberibacter ◽  
Effective Program ◽  
Liberibacter Asiaticus ◽  
Root Tissues

Huanglongbing (HLB), or citrus greening, is the most destructive disease to the citrus industry. In Florida, it is caused by the bacterium, Candidatus Liberibacter asiaticus (CLas) and is transmitted by the Asian citrus psyllid, Diaphorina citri. Recent studies suggested that antibiotics could inhibit the growth of the CLas pathogen in planta. In the current study, we investigated the uptake and translocation of oxytetracycline and streptomycin in citrus seedlings. Oxytetracycline and streptomycin were delivered via root and stem and their level in various tissues was monitored using enzyme-linked immunosorbent assay (ELISA). Oxytetracycline and streptomycin were detected in the leaves, xylem, phloem, and root after root drench and stem delivery. High levels of antibiotics were detected in the roots after root drench, whereas high levels of antibiotics were detected in the canopy after stem delivery. The level of oxytetracycline detected in the phloem, xylem, and leaves after root drench was higher than that of streptomycin. Whereas the level of streptomycin in root was higher than that of oxytetracycline, indicating that streptomycin was bound to the xylem tissues. Oxytetracycline and streptomycin were detected in the phloem, xylem, leaves, and root tissues thirty-five days after the root incubation in 200 µg·mL−1 solution. These results demonstrated that oxytetracycline and streptomycin were relatively stable and could inhibit CLas growth for a couple of months in citrus trees. Observations reported in this study regarding the distribution and stability of oxytetracycline and streptomycin in citrus plants could be useful for designing an effective program for the control of HLB disease using antibiotics.

scholarly journals Transcriptomic and Metabolomic Analyses of Diaphorina citri Kuwayama Infected and Non-infected With Candidatus Liberibacter Asiaticus

2021 ◽  
Vol 11 ◽  
Author(s):  
Kai Liu ◽  
Jiawei He ◽  
Ziying Guan ◽  
Mingzhao Zhong ◽  
Rui Pang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Host Plants ◽  
Integrated Analysis ◽  
Asian Citrus Psyllid ◽  
Insect Vector ◽  
Diaphorina Citri ◽  
Liquid Chromatography Mass Spectrometry ◽  
Candidatus Liberibacter Asiaticus ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

The Asian citrus psyllid Diaphorina citri is the transmission vector of Huanglongbing (HLB), a devastating disease of citrus plants. The bacterium “Candidatus Liberibacter asiaticus” (CLas) associated with HLB is transmitted between host plants by D. citri in a circulative manner. Understanding the interaction between CLas and its insect vector is key for protecting citrus cultivation from HLB damage. Here, we used RNA sequencing and liquid chromatography-mass spectrometry (LC-MS) to analyze the transcriptome and metabolome of D. citri interacting with CLas. We identified 662 upregulated and 532 downregulated genes in CLas-infected insects. These genes were enriched in pathways involving carbohydrate metabolism, the insects’ immune system, and metabolism of cofactors and vitamins. We also detected 105 differential metabolites between CLas-infected and non-infected insects, including multiple nucleosides and lipid-related molecules. The integrated analysis revealed nine pathways—including those of the glycine, serine, threonine, and purine metabolism—affected by the differentially expressed genes from both groups. The network for these pathways was subsequently constructed. Our results thus provide insights regarding the cross-talk between the transcriptomic and metabolomic changes in D. citri in response to CLas infection, as well as information on the pathways and genes/metabolites related to the CLas–D. citri interaction.

scholarly journals Susceptibility of Sixteen Citrus Genotypes to ‘Candidatus Liberibacter asiaticus’

Plant Disease ◽  
2016 ◽  
Vol 100 (6) ◽  
pp. 1080-1086 ◽  
Author(s):  
Greg McCollum ◽  
Mark Hilf ◽  
Mike Irey ◽  
Weiqi Luo ◽  
Tim Gottwald
Keyword(s):  
The United States ◽  
Insect Vector ◽  
Candidatus Liberibacter Asiaticus ◽  
Citrus Germplasm ◽  
Citrus Production ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus ◽  
Florida Citrus ◽  
Citrus Macrophylla ◽  
Free Ranging

Huanglongbing (HLB) disease is the most serious threat to citrus production worldwide and, in the last decade, has devastated the Florida citrus industry. In the United States, HLB is associated with the phloem-limited α-proteobacterium ‘Candidatus Liberibacter asiaticus’ and its insect vector, the Asian citrus psyllid (ACP; Diaphorina citri). Significant effort is being put forth to develop novel citrus germplasm that has a lower propensity to succumb to HLB than do currently available varieties. Effective methods of screening citrus germplasm for susceptibility to HLB are essential. In this study, we exposed small, grafted trees of 16 citrus types to free-ranging ACP vectors and ‘Ca. L. asiaticus’ inoculum in the greenhouse. During 45 weeks of exposure to ACP, the cumulative incidence of ‘Ca. L. asiaticus’ infection was 70%. Trees of Citrus macrophylla and C. medica were most susceptible to ‘Ca. L. asiaticus’, with 100% infection by the end of the test period in three trials, while the complex genetic hybrids ‘US 1-4-59’ and ‘Fallglo’ consistently were least susceptible, with approximately 30% infection. Results obtained in this greenhouse experiment showed good agreement with trends observed in the orchard, supporting the validity of our approach for screening citrus germplasm for susceptibility to HLB.

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