scholarly journals ‘Candidatus Liberibacter asiaticus’ and Its Vector, Diaphorina citri, Augment the Tricarboxylic Acid Cycle of Their Host via the γ-Aminobutyric Acid Shunt and Polyamines Pathway

2019 ◽  
Vol 32 (4) ◽  
pp. 413-427 ◽  
Author(s):  
Yasser Nehela ◽  
Nabil Killiny
Keyword(s):  
Tca Cycle ◽  
Tricarboxylic Acid ◽  
Aminobutyric Acid ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Gaba Shunt ◽  
The Tca Cycle ◽  
Candidatus Liberibacter ◽  
Nutritional Needs ◽  
Liberibacter Asiaticus

Huanglongbing (HLB), a destructive citrus disease, is associated with ‘Candidatus Liberibacter asiaticus’, which is transmitted by the Asian citrus psyllid Diaphorina citri. Both ‘Ca. L. asiaticus’ and its vector manipulate the host metabolism for their benefit, to meet their nutritional needs and neutralize the host defense responses. We used a targeted gas chromatography-mass spectrometry–based method to explore the connection between the tricarboxylic acid (TCA) cycle, γ-aminobutyric acid (GABA) shunt, and polyamines (PAs) pathways in citrus. ‘Ca. L. asiaticus’ and D. citri accelerated the conversion of α-ketoglutarate to glutamate, then to GABA, causing an accumulation of GABA in the cytosol. In silico analysis showed that the citrus genome possesses a putative GABA permease that connects the GABA shunt with the TCA cycle and supports the accumulation of succinate, fumarate, and citrate. Additionally, the PAs biosynthetic pathway might be connected directly to the TCA cycle, through the production of fumarate, or indirectly, via enhancement of GABA shunt. Taken together, we suggest that GABA shunt and PAs pathways are alternative pathways that contribute to the flux toward succinate rather than an intact TCA cycle in citrus. Both ‘Ca. L. asiaticus’ and its vector enhance these pathways. This study provides more insights into citrus responses to the HLB pathosystem and could be a further step toward clues for understanding the nutritional needs of ‘Ca. L. asiaticus’, which could help in culturing ‘Ca. L. asiaticus’.

scholarly journals Not Just a Cycle: Three gab genes Enable the Non-cyclic Flux Toward Succinate Via GABA Shunt in ‘Candidatus Liberibacter asiaticus’-infected citrus

2021 ◽  
Author(s):  
Yasser Nehela ◽  
Nabil Killiny
Keyword(s):  
Tca Cycle ◽  
Defense Responses ◽  
Candidatus Liberibacter Asiaticus ◽  
Amino Acid Permease ◽  
Phytopathogenic Bacterium ◽  
Gaba Shunt ◽  
Semialdehyde Dehydrogenase ◽  
The Tca Cycle ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

Although the mitochondria retain all required enzymes for an intact tricarboxylic acid (TCA) cycle, plants might shift the cyclic flux from the TCA cycle to an alternative non-cyclic pathway via γ-aminobutyric acid (GABA) shunt under specific physiological conditions. We hypothesize that several genes may ease this non-cyclic flux and contribute to the citrus response to the phytopathogenic bacterium ‘Candidatus Liberibacter asiaticus’, the causal agent of Huanglongbing in citrus. To test this hypothesis, we used multi-omics techniques (metabolomics, fluxomics, and transcriptomics) to investigate the potential role(s) of putative gab homologies from Valencia sweet orange (Citrus sinensis). Our findings showed that ‘Ca. L. asiaticus’ significantly increased the endogenous GABA and succinate content but decreased ketoglutarate in infected citrus plants. Citrus genome harbors three putative gab genes including amino-acid permease (aka GABA permease; CsgabP), GABA transaminase (CsgabT), and succinate-semialdehyde dehydrogenase (aka GABA dehydrogenase; CsgabD). The transcript levels of CsgabP, CsgabT, and CsgabD were upregulated in citrus leaves upon the infection with ‘Ca. L. asiaticus’ and after the exogenous application of GABA or deuterium-labeled GABA isotope (GABA-D6). Moreover, our finding showed that exogenously applied GABA is quickly converted to succinate and fed into the TCA cycle. Likewise, the fluxomics study showed that GABA-D6 is rapidly metabolized to succinate-D4. Our work proved that GABA shunt and three predicated gab genes from citrus, support the upstream non-cyclic flux toward succinate rather than an intact TCA cycle and contribute to citrus defense responses to ‘Ca. L. asiaticus’.

Generous hosts: Growth of ‘Candidatus Liberibacter asiaticus’ in Madagascar periwinkle (Catharanthus roseus) highlights its nutritional needs

2021 ◽  
Author(s):  
Nabil Killiny
Keyword(s):  
Organic Acids ◽  
Catharanthus Roseus ◽  
Bacterial Growth ◽  
Tca Cycle ◽  
Candidatus Liberibacter Asiaticus ◽  
Madagascar Periwinkle ◽  
Phloem Sap ◽  
The Tca Cycle ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

‛Candidatus Liberibacter asiaticus’, the putative causal agent of citrus greening is not available in pure culture yet. In addition to trees of citrus and citrus relatives, ‛Ca. L. asiaticus’ can grow in Madagascar periwinkle (Catharanthus roseus). Using GC-MS, we compared the phloem sap composition in sweet orange ‛Valencia’ (Citrus sinensis) and periwinkle plants after the infection with ‛Ca. L. asiaticus’. Interestingly, in contrast to our previous studies of total leaf metabolites, we found that, compared to uninfected phloem sap, we found that the organic acids implicated in the TCA cycle including citrate, isocitrate, succinate, fumarate, and malate were reduced significantly in the infected phloem saps of both species. As a result of the reduction of organic acids content, the pH of infected phloem saps was increased. We hypothesize that the bacterial growth induces the mitochondrial TCA cycle in parenchyma cells to produce more of these compounds to be used as a bacterial carbon source. Once these compounds reach a low level in the phloem sap, the bacterium may send a signal, yet to be identified, to initiate a feedback loop to further induce the TCA cycle. Phloem blockage might be another reason behind the reduced the translocation of TCA cycle intermediates within the phloem. The net result, localized availability of organic acids likely benefits the bacterial growth and may explain the unequal distribution of ‘Ca L. asiaticus’ within infected trees. These findings may help in designing media for the pure culturing of ‛Ca. L. asiaticus’.

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.

A Review of Huanglongbing (Citrus Greening) Management in Citrus Using Nutritional Approaches in China

2011 ◽  
Vol 12 (1) ◽  
pp. 24 ◽  
Author(s):  
Yulu Xia ◽  
Gecheng Ouyang ◽  
Ronald A. Sequeira ◽  
Yu Takeuchi ◽  
Ignacio Baez ◽  
...  
Keyword(s):  
Nutrient Management ◽  
Field Survey ◽  
Cultural Practices ◽  
Sweet Orange ◽  
Asian Citrus Psyllid ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Candidatus Liberibacter ◽  
Citrus Maxima ◽  
Liberibacter Asiaticus

The Asian form of huanglongbing (HLB) is caused by ‘Candidatus Liberibacter asiaticus (Las),’ a phloem-limited bacterium transmitted by the Asian citrus psyllid, Diaphorina citri Kuwayama. Nutrient management, together with other cultural practices such as pruning and irrigation, for mitigation of the disease has been practiced in China for many years. Our literature review, field survey, and interviews with Chinese scientists and growers indicate that these cultural practices were generally ineffective for the disease management. However, a nutritional approach in conjunction with other cultural practices such as irrigation can maintain grove productivity for a certain time depending on the type of citrus species/cultivars, the age of the trees, the propagation method of the plants, the Asian citrus psyllid (ACP) (Diaphorina citri Kuwayama) population, and other factors. Symptomatic mature pommelo (Citrus maxima Merr) and sweet orange (C. sinensis L. Osbeck) plants can commonly survive and maintain a certain level of productivity for an additional 4 to 5 years, even longer assuming vigorous ACP control. Accepted for publication 27 June 2011. Published 3 October 2011.

scholarly journals Tracing Penicillin Movement in Citrus Plants Using Fluorescence-Labeled Penicillin

Antibiotics ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 262
Author(s):  
Nabil Killiny ◽  
Pedro Gonzalez-Blanco ◽  
Yulica Santos-Ortega ◽  
Fuad Al-Rimawi ◽  
Amit Levy ◽  
...  
Keyword(s):  
Vascular Tissue ◽  
Vascular System ◽  
Citrus Greening ◽  
Application Site ◽  
Asian Citrus Psyllid ◽  
Diaphorina Citri ◽  
Candidatus Liberibacter Asiaticus ◽  
Citrus Industry ◽  
Candidatus Liberibacter ◽  
Liberibacter Asiaticus

Huánglóngbìng (HLB), citrus greening, is one of the most destructive diseases of citrus plants worldwide. In North America, HLB is caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus and is transmitted by the Asian citrus psyllid, Diaphorina citri. No cure exists at present, and the use of antibiotics for the control of HLB has gained interest due to the significant losses to the citrus industry. Because of unsatisfactory results when using foliar applications of antibiotics, concerns were raised regarding the uptake and translocation of these materials within trees. We, therefore, investigated a method that allows us to study the movement of antibiotic materials in citrus plants. Herein, we utilized a fluorescence-labeled penicillin, BOCILLIN™ FL-Penicillin (FL-penicillin), to study the uptake and translocation of penicillin in citrus plants. FL-penicillin was applied by puncture to the stem of young citrus seedlings and was traced by using fluorescence microscopy. After application, we detected FL-penicillin in the leaves and in the stem xylem and phloem tissues above and below the application site in both intact and partially bark-girdled citrus seedlings, indicating that it is easily taken up and transported through the plant vascular system. In addition, we detected FL-penicillin in the gut of D. citri, which were allowed to feed on the treated plants, suggesting translocation of this molecule into the vascular tissue. We propose that the use of fluorescent-labeled molecules could be an effective tool for understanding the uptake and translocation of antibiotics and other macromolecules in plants and insects.

Sign in / Sign up

Export Citation Format

Share Document