James C. K. Ng
University of California, Riverside
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Molecular Plant Pathology | 2004
James C. K. Ng; Keith L. Perry
SUMMARY Aphids are the most common vector of plant viruses. Mechanisms of transmission are best understood by considering the routes of virus movement in the aphid (circulative versus non-circulative) and the sites of retention or target tissues (e.g. stylets, salivary glands). Capsid proteins are a primary, but not necessarily sole, viral determinant of transmission. A summary is presented of the taxonomic affiliations of the aphid transmitted viruses, including 8 families, 18 genera, and taxonomically unassigned viruses.
Current Microbiology | 2003
MyLo Ly Thao; Linda Baumann; Justin M. Hess; Bryce W. Falk; James C. K. Ng; Penny J. Gullan; Paul Baumann
On the basis of 16S–23S ribosomal DNA analyses, the whitefly Bemisia tabaci (Sternorrhyncha, Aleyrodidae) and the eriococcid Eriococcus spurius (Sternorrhyncha, Eriococcidae) were each found to harbor novel related chlamydial species within the family Simkaniaceae. The generic designation Fritschea gen. nov. is proposed to accommodate the two species, F. bemisiae sp. nov. and F. eriococci sp. nov. The finding of chlamydial 16S–23S ribosomal DNA in B. tabaci is consistent with a previous electron microscopy study which found that bacteriocytes of this species contain structures that we consider to resemble the elementary and reticulate bodies of chlamydia (Costa HS, Westcot DM, Ullman DE, Rosell R, Brown JK, Johnson MW. Protoplasma 189:194–202, 1995). The cloning and sequencing of a 16.6 kilobase DNA fragment from F. bemisiae indicated that it contains six genes encoding for proteins similar to those found in other species of chlamydia. These results extend the range of organisms that harbor chlamydia.
Proceedings of the National Academy of Sciences of the United States of America | 2011
Angel Y.S. Chen; G. P. Walker; David Carter; James C. K. Ng
Numerous pathogens of humans, animals, and plants are transmitted by specific arthropod vectors. However, understanding the mechanisms governing these pathogen–vector interactions is hampered, in part, by the lack of easy-to-use analytical tools. We investigated whitefly transmission of Lettuce infectious yellows virus (LIYV) by using a unique immunofluorescent localization approach in which we fed virions or recombinant virus capsid components to whiteflies, followed by feeding them antibodies to the virions or capsid components, respectively. Fluorescent signals, indicating the retention of virions, were localized in the anterior foregut or cibarium of a whitefly vector biotype but not within those of a whitefly nonvector biotype. Retention of virions in these locations strongly corresponded with the whitefly vector transmission of LIYV. When four recombinant LIYV capsid components were individually fed to whitefly vectors, significantly more whiteflies retained the recombinant minor coat protein (CPm). As demonstrated previously and in the present study, whitefly vectors failed to transmit virions preincubated with anti-CPm antibodies but transmitted virions preincubated with antibodies recognizing the major coat protein (CP). Correspondingly, the number of insects that specifically retained virions preincubated with anti-CPm antibodies were significantly reduced compared with those that specifically retained virions preincubated with anti-CP antibodies. Notably, a transmission-defective CPm mutant was deficient in specific virion retention, whereas the CPm-restored virus showed WT levels of specific virion retention and transmission. These data provide strong evidence that transmission of LIYV is determined by a CPm-mediated virion retention mechanism in the anterior foregut or cibarium of whitefly vectors.
Journal of Virology | 2010
Lucy R. Stewart; Vicente Medina; Tongyan Tian; Massimo Turina; Bryce W. Falk; James C. K. Ng
ABSTRACT The Lettuce infectious yellows virus (LIYV) RNA 2 mutant p1-5b was previously isolated from Bemisia tabaci-transmitted virus maintained in Chenopodium murale plants. p1-5b RNA 2 contains a single-nucleotide deletion in the minor coat protein (CPm) open reading frame (ORF) that is predicted to result in a frameshift and premature termination of the protein. Using the recently developed agroinoculation system for LIYV, we tested RNA 2 containing the p1-5b CPm mutant genotype (agro-pR6-5b) in Nicotiana benthamiana plants. We showed that plant infection triggered by agro-pR6-5b spread systemically and resulted in the formation of virions similar to those produced in p1-5b-inoculated protoplasts. However, virions derived from these mutant CPm genotypes were not transmitted by whiteflies, even though virion concentrations were above the typical transmission thresholds. In contrast, and as demonstrated for the first time, an engineered restoration mutant (agro-pR6-5bM1) was capable of both systemic movement in plants and whitefly transmission. These results provide strong molecular evidence that the full-length LIYV-encoded CPm is dispensable for systemic plant movement but is required for whitefly transmission.
Current Microbiology | 2004
Linda Baumann; MyLo Ly Thao; C. Joel Funk; Bryce W. Falk; James C. K. Ng; Paul Baumann
The whitefly Bemisia tabaci contains a primary prokaryotic endosymbiont housed within specialized cells in the body cavity. Two DNA fragments from the endosymbiont, totaling 33.3 kilobases, were cloned and sequenced. In total, 37 genes were detected and included the ribosomal RNA operon and genes for ribosomal RNA proteins. The guanine plus cytosine of the DNA was 30.2 mol%, different from that of endosymbionts of other plant sap-sucking insects.
Current Opinion in Virology | 2015
James C. K. Ng; Jaclyn S. Zhou
The non-circulative, semi-persistent (NCSP) mode of insect vector-mediated plant virus transmission is shaped by biological, molecular and mechanical interactions that take place across a continuum of processes involved in virion acquisition, retention and inoculation. Our understanding of the interactive roles of virus, insect vector, and plant associated with NCSP transmission is still evolving. Mechanisms exist that determine where and how virion acquisition (from the plant) and retention (in the insect vector) are achieved, with both processes being mediated by strategies involving viral capsid proteins, in some cases aided by non-capsid proteins. By contrast, mechanisms underlying virion inoculation (to the plant) remain poorly understood. Here, we review the established paradigms as well as fresh perspectives on the mechanisms of NCSP transmission.
Analytical Methods | 2013
Nicha Chartuprayoon; Youngwoo Rheem; James C. K. Ng; Jin Nam; Wilfred Chen; Nosang V. Myung
Label-free chemiresistive sensors based on a polypyrrole (PPy) nanoribbon (width: 500 nm, thickness: 25–100 nm) were batch-fabricated by a lithographically patterned nanowire electrodeposition (LPNE) technique. A plant pathogen specific antibody was covalently conjugated on the surface of the structure via N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDC)/N-hydrosuccinimide (NHS) crosslinking. The sensing performance was investigated by the detection of cucumber mosaic virus (CMV). The sensitivity of the nano-immunosensors was enhanced by reducing the electrical conductivity from 1 to 0.005 S cm−1 or by decreasing the thickness of the nanoribbon from 100 nm to 25 nm. The reduction in the ionic strength of the pH buffer solutions (i.e., 10 mM PBS to 10 mM PB) also enhanced the sensitivity. However, the reliability and reproducibility of the sensors were significantly reduced by the buffer change. The optimum sensor showed excellent sensitivity with a low and upper detection limit of 10 ng ml−1 and 100 μg ml−1, respectively, which is much lower than the low detection limit of traditional enzyme-linked immunosorbent assays (ELISAs) (i.e., 3 μg ml−1).
Frontiers in Microbiology | 2013
James C. K. Ng
Successful vector-mediated plant virus transmission entails an intricate but poorly understood interplay of interactions among virus, vector, and plant. The complexity of interactions requires continually improving/evaluating tools and methods for investigating the determinants that are central to mediating virus transmission. A recent study using an organic fluorophore (Alexa Fluor)-based immunofluorescent localization assay demonstrated that specific retention of Lettuce infectious yellows virus (LIYV) virions in the anterior foregut or cibarium of its whitefly vector is required for virus transmission. Continuous exposure of organic fluorophore to high excitation light intensity can result in diminished or loss of signals, potentially confounding the identification of important interactions associated with virus transmission. This limitation can be circumvented by incorporation of photostable fluorescent nanocrystals, such as quantum dots (QDs), into the assay. We have developed and evaluated a QD-immunofluorescent labeling method for the in vitro and in situ localization of LIYV virions based on the recognition specificity of streptavidin-conjugated QD605 (S-QD605) for biotin-conjugated anti-LIYV IgG (B-αIgG). IgG biotinylation was verified in a blot overlay assay by probing SDS-PAGE separated B-αIgG with S-QD605. Immunoblot analyses of LIYV using B-αIgG and S-QD605 resulted in a virus detection limit comparable to that of DAS-ELISA. In membrane feeding experiments, QD signals were observed in the anterior foregut or cibarium of virion-fed whitefly vectors but absent in those of virion-fed whitefly non-vectors. Specific virion retention in whitefly vectors corresponded with successful virus transmission. A fluorescence photobleaching assay of viruliferous whiteflies fed B-αIgG and S-QD605 vs. those fed anti-LIYV IgG and Alexa Fluor 488-conjugated IgG revealed that QD signal was stable and deteriorated approx. seven- to eight-fold slower than that of Alexa Fluor.
Insect Science | 2017
Beatriz Dáder; Christiane Then; Edwige Berthelot; Marie Ducousso; James C. K. Ng; Martin Drucker
By serving as vectors of transmission, insects play a key role in the infection cycle of many plant viruses. Viruses use sophisticated transmission strategies to overcome the spatial barrier separating plants and the impediment imposed by the plant cell wall. Interactions among insect vectors, viruses, and host plants mediate transmission by integrating all organizational levels, from molecules to populations. Best‐examined on the molecular scale are two basic transmission modes wherein virus–vector interactions have been well characterized. Whereas association of virus particles with specific sites in the vectors mouthparts or in alimentary tract regions immediately posterior to them is required for noncirculative transmission, the cycle of particles through the vector body is necessary for circulative transmission. Virus transmission is also determined by interactions that are associated with changes in vector feeding behaviors and with alterations in plant hosts morphology and/or metabolism that favor the attraction or deterrence of vectors. A recent concept in virus–host–vector interactions proposes that when vectors land on infected plants, vector elicitors and effectors “inform” the plants of the confluence of interacting entities and trigger signaling pathways and plant defenses. Simultaneously, the plant responses may also influence virus acquisition and inoculation by vectors. Overall, a picture is emerging where transmission depends on multilayered virus–vector–host interactions that define the route of a virus through the vector, and on the manipulation of the host and the vector. These interactions guarantee virus propagation until one or more of the interactants undergo changes through evolution or are halted by environmental interventions.
Virus Research | 2011
James C. K. Ng; Angel Y.S. Chen
Viruses in the genus Crinivirus infect diverse plant species and are transmitted by specific whitefly vectors, but the basis for vector specific transmission remains poorly understood. Here, we demonstrated that purified virion preparations of Lettuce chlorosis virus (LCV) contained filamentous particles that were consistently transmitted to plants by whiteflies (Bemisia tabaci biotypes A and B) following membrane feeding, suggesting that the preparations contained biologically active virions with all the components essential for specific vector transmission. We also demonstrated in sequential membrane feeding experiments that B. tabaci biotype A pre-fed with high concentrations of Lettuce infectious yellows virus (LIYV) virions followed by decreasing concentrations of LCV virions either abolished or interfered with the transmission of the latter. However, in the reverse treatment, an abolishment/interference in transmission of LIYV was not observed. These results suggest that both viruses share a common transmission pathway in B. tabaci biotype A, and factors other than virion quality and quantity may additionally influence their transmission. To begin investigating the viral determinants that are involved in mediating the whitefly transmission of LCV, virions were analyzed by Western immunoblotting. Our results showed that virions were positively identified by antisera produced against three E. coli expressed recombinant LCV capsid proteins--the major coat protein [CP], minor CP [CPm], and P60.