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Title: Phytoreovirus-like sequences isolated from salivary glands of the glassy-winged sharpshooter Homolodisca vitripennis (Hemiptera: Cicadellidae)

Abstract

The salivary glands of the Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis Germar 1821, (syn. H. coagulata, Hemiptera: Cicadellidae) were collected and used to produce a cDNA library. Examination by BLASTX analyses identified 2 viral sequences, one a 610-base pair fragment and a second 839-base pair fragment, both of which had significant homology to viruses within the genus Phytoreovirus. Resequencing of the fragments confirmed sequence validities. These sequences were used for in silico protein translation and BLASTP analysis confirming the established homology. While the GWSS is the primary vector of Pierce's disease of grapes, this is the first report that GWSS may be a vector of a phytoreoviruses. Phylogenetic and homology comparisons with BLASTX, BLASTP, and PAUP analyses indicated that the viral sequences isolated from GWSS were closely related to the viruses in the Family Reoviridae, Genus Phytoreovirus, specifically Rice Dwarf Phytoreovirus (RDV). RDV is the only plant reovirus that is not limited to the phloem. Phytoreoviruses are transmitted in a propagative manner by cicadellid leafhoppers (Hemiptera: Cicadellidae), which acquire and transmit them during feeding. Phytoreoviruses have been reported from Agallian, Agalliopsis, Nephotettix, and Recilia, genera of leafhoppers, with evidence for transovarial transmission. The GWSS, although considered to feed primarily from themore » xylem, ingests from other plant tissues, such as the phloem and mesophyll during probing similar to other leafhoppers. The feeding behavior and wide host range of the GWSS provides an overlapping condition for these two organisms, leafhopper and virus. GWSS will feed from grasses as a transitory host, and on herbaceous and woody plants as primary hosts, which may favor the acquisition and transmission of Phytoreovirus by this leafhopper. Monitoring for an increase of Phytoreovirus spread in graminaceous crops that are in proximity to vineyards or tree crop orchards, where GWSS occurs, such as in southern California, will provide a better understanding of the potential role of the GWSS as a disease vector in the spread of phytoreoviruses and other plant pathogens. The sequences have been deposited in NCBI database under the accession numbers (EF058280) for GWSS-V1, WHSg013C11 and (EF058281) for GWSS-V2, WHSg024H02. (author) [Spanish] Dos fragmentos de 610 y 839 pares de bases fueron aislados apartir de una genoteca de expresion derivada de las glandulas salivales del cucarron de las alas cristalinas (GWSS), Homalodisca vitripennis, Germar 1821 (syn. H. coagulata) el cual es vector de la enfermedad de Pierce de las uvas. Los resultados de alineamiento utilizando BLASTX, BLASTP y el analisis filogenetico utilizando PAUP indicaron que los fragmentos de DNA estaban relacionado de manera mas cercana a viruses en la familia Reoviridae, genero Phytoreovirus, y especificamente a los virus del enanismo del arroz (RDV) y al virus del tumor de las grietas (WTV). El cucarron de las alas cristalinas es un saltahoja que se alimenta no solo del xilema sino tambien del floema y del mesofilo. Saltahojas del genero Agallian, son los principales vectores de WTV, el cual infecta el floema de plantas dicotiledoneas tumoraciones en las hojas y en las raices. WTV es transmitido por saltahojas y es el unico reovirus que es capaz de infectar tanto tejidos del xilema como del floema. El comportamiento alimentario del GWSS y su amplio rango de hospederos que incluye pastos y plantas herbaceas y lenosas podria proveer la interaccion entre estos dos organismos facilitando la adquisicion y tranmision de fitoreovirus por el GWSS. Un adecuado monitoreo de el incremento en la expansion de reoviruses en cultivos de gramineas asociados al los vinedos en donde GWSS ocurre en regiones tales como el sur de California, y en general el sur de los Estados Unidos, podria proveer un mejor entendimiento del papel del GWSS como vector de fitoreovirus y otros patogenos de plantas. Las sequencias se depositaron en la base de datos NCBI con los siguientes numeros de identificacion: (EF058280) para GWSS-V1, WHSg013C11 y (EF058281) para GWSSV2, WHSg024H02. (author)« less

Authors:
;  [1];  [2]
  1. Subtropical Insect Research Unit, United States Horticultural Research Lab, United States Department Agriculture, ARS, Fort Pierce, FL 34945 (United States). E-mail: Whunter@ushrl.ars.usda.gov
  2. University of Florida, Institute of Food and Agricultural Sciences, Indian River Research and Education Center, Fort Pierce, FL 34945 (United States)
Publication Date:
OSTI Identifier:
20942929
Resource Type:
Journal Article
Resource Relation:
Journal Name: Florida Entomologist; Journal Volume: 90; Journal Issue: 1; Other Information: Country of input: International Atomic Energy Agency (IAEA); DOI: 10.1653/0015-4040(2007)90[196:PSIFSG]2.0.CO;2; Spanish abstract translation provided by the authors; Refs, 4 figs, 2 tabs
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CALIFORNIA; CROPS; DISEASE VECTORS; FEEDING; GRAPES; HEMIPTERA; ONCOGENIC VIRUSES; PATHOGENS; PLANT DISEASES; PLANT TISSUES; PROTEINS; RICE; SALIVARY GLANDS; TREES

Citation Formats

Katsar, C.S., Hunter, W.B., and Sinisterra, X.H. Phytoreovirus-like sequences isolated from salivary glands of the glassy-winged sharpshooter Homolodisca vitripennis (Hemiptera: Cicadellidae). United States: N. p., 2007. Web. doi:10.1653/0015-4040(2007)90[196:PSIFSG]2.0.CO;2; SPANISH ABSTRACT TRANSLATION PROVIDED BY THE AUTHORS; REFS, 4 FIGS, 2 TABS.
Katsar, C.S., Hunter, W.B., & Sinisterra, X.H. Phytoreovirus-like sequences isolated from salivary glands of the glassy-winged sharpshooter Homolodisca vitripennis (Hemiptera: Cicadellidae). United States. doi:10.1653/0015-4040(2007)90[196:PSIFSG]2.0.CO;2; SPANISH ABSTRACT TRANSLATION PROVIDED BY THE AUTHORS; REFS, 4 FIGS, 2 TABS.
Katsar, C.S., Hunter, W.B., and Sinisterra, X.H. Thu . "Phytoreovirus-like sequences isolated from salivary glands of the glassy-winged sharpshooter Homolodisca vitripennis (Hemiptera: Cicadellidae)". United States. doi:10.1653/0015-4040(2007)90[196:PSIFSG]2.0.CO;2; SPANISH ABSTRACT TRANSLATION PROVIDED BY THE AUTHORS; REFS, 4 FIGS, 2 TABS.
@article{osti_20942929,
title = {Phytoreovirus-like sequences isolated from salivary glands of the glassy-winged sharpshooter Homolodisca vitripennis (Hemiptera: Cicadellidae)},
author = {Katsar, C.S. and Hunter, W.B. and Sinisterra, X.H.},
abstractNote = {The salivary glands of the Glassy-winged sharpshooter (GWSS), Homalodisca vitripennis Germar 1821, (syn. H. coagulata, Hemiptera: Cicadellidae) were collected and used to produce a cDNA library. Examination by BLASTX analyses identified 2 viral sequences, one a 610-base pair fragment and a second 839-base pair fragment, both of which had significant homology to viruses within the genus Phytoreovirus. Resequencing of the fragments confirmed sequence validities. These sequences were used for in silico protein translation and BLASTP analysis confirming the established homology. While the GWSS is the primary vector of Pierce's disease of grapes, this is the first report that GWSS may be a vector of a phytoreoviruses. Phylogenetic and homology comparisons with BLASTX, BLASTP, and PAUP analyses indicated that the viral sequences isolated from GWSS were closely related to the viruses in the Family Reoviridae, Genus Phytoreovirus, specifically Rice Dwarf Phytoreovirus (RDV). RDV is the only plant reovirus that is not limited to the phloem. Phytoreoviruses are transmitted in a propagative manner by cicadellid leafhoppers (Hemiptera: Cicadellidae), which acquire and transmit them during feeding. Phytoreoviruses have been reported from Agallian, Agalliopsis, Nephotettix, and Recilia, genera of leafhoppers, with evidence for transovarial transmission. The GWSS, although considered to feed primarily from the xylem, ingests from other plant tissues, such as the phloem and mesophyll during probing similar to other leafhoppers. The feeding behavior and wide host range of the GWSS provides an overlapping condition for these two organisms, leafhopper and virus. GWSS will feed from grasses as a transitory host, and on herbaceous and woody plants as primary hosts, which may favor the acquisition and transmission of Phytoreovirus by this leafhopper. Monitoring for an increase of Phytoreovirus spread in graminaceous crops that are in proximity to vineyards or tree crop orchards, where GWSS occurs, such as in southern California, will provide a better understanding of the potential role of the GWSS as a disease vector in the spread of phytoreoviruses and other plant pathogens. The sequences have been deposited in NCBI database under the accession numbers (EF058280) for GWSS-V1, WHSg013C11 and (EF058281) for GWSS-V2, WHSg024H02. (author) [Spanish] Dos fragmentos de 610 y 839 pares de bases fueron aislados apartir de una genoteca de expresion derivada de las glandulas salivales del cucarron de las alas cristalinas (GWSS), Homalodisca vitripennis, Germar 1821 (syn. H. coagulata) el cual es vector de la enfermedad de Pierce de las uvas. Los resultados de alineamiento utilizando BLASTX, BLASTP y el analisis filogenetico utilizando PAUP indicaron que los fragmentos de DNA estaban relacionado de manera mas cercana a viruses en la familia Reoviridae, genero Phytoreovirus, y especificamente a los virus del enanismo del arroz (RDV) y al virus del tumor de las grietas (WTV). El cucarron de las alas cristalinas es un saltahoja que se alimenta no solo del xilema sino tambien del floema y del mesofilo. Saltahojas del genero Agallian, son los principales vectores de WTV, el cual infecta el floema de plantas dicotiledoneas tumoraciones en las hojas y en las raices. WTV es transmitido por saltahojas y es el unico reovirus que es capaz de infectar tanto tejidos del xilema como del floema. El comportamiento alimentario del GWSS y su amplio rango de hospederos que incluye pastos y plantas herbaceas y lenosas podria proveer la interaccion entre estos dos organismos facilitando la adquisicion y tranmision de fitoreovirus por el GWSS. Un adecuado monitoreo de el incremento en la expansion de reoviruses en cultivos de gramineas asociados al los vinedos en donde GWSS ocurre en regiones tales como el sur de California, y en general el sur de los Estados Unidos, podria proveer un mejor entendimiento del papel del GWSS como vector de fitoreovirus y otros patogenos de plantas. Las sequencias se depositaron en la base de datos NCBI con los siguientes numeros de identificacion: (EF058280) para GWSS-V1, WHSg013C11 y (EF058281) para GWSSV2, WHSg024H02. (author)},
doi = {10.1653/0015-4040(2007)90[196:PSIFSG]2.0.CO;2; SPANISH ABSTRACT TRANSLATION PROVIDED BY THE AUTHORS; REFS, 4 FIGS, 2 TABS},
journal = {Florida Entomologist},
number = 1,
volume = 90,
place = {United States},
year = {Thu Mar 15 00:00:00 EDT 2007},
month = {Thu Mar 15 00:00:00 EDT 2007}
}
  • A new virus species of the genus Phytoreovirus was isolated from glassy-winged sharpshooter (GWSS), Homalodisca vitripennis Germar (Hemiptera: Cicadellidae), in California and designated here as Homalodisca vitripennis reovirus (HoVRV). Extraction of nucleic acid from GWSS adults collected from three Californian populations revealed an array of double-stranded (ds) RNA species that was soluble in 2 M LiCl and resistant to degradation upon exposure to S1 nuclease and DNase. Analysis of nucleic acid samples from single GWSS adults indicated that HoVRV dsRNA accumulated to high titer in individual insects. Double-shelled isometric virus particles purified from GWSS adults resembled those observed in thinmore » sections of GWSS salivary glands by transmission electron microscopy. Purified HoVRV virions contained 12 dsRNA segments that, based on complete nucleotide sequences, ranged in size from 4475 to 1040 bp. Sequence comparisons indicated that the HoVRV dsRNA segments were most closely related (58.5 to 43.7% nt sequence identity) to the corresponding genome segments of Rice dwarf virus (RDV). Each HoVRV dsRNA segment encoded a single open reading frame (> 300 nts) except for segment 11, which appears to be dicistronic. Terminal nucleotide sequences of HoVRV positive-sense RNAs were similar to other phytoreoviruses (GGCG or GGCA at the 5'-end and UGAU or CGAU at the 3'-end) with adjacent imperfect inverted repeats potentially able to base pair. Phylogenetic analyses of the RNA-directed RNA polymerase (encoded by segment 1) and the outer capsid protein (encoded by segment 8) confirmed placement of HoVRV as a species of the genus Phytoreovirus sharing a most recent common ancestor with RDV. Reverse transcription-polymerase chain reaction assays revealed that HoVRV infection of GWSS in California was common and that the virus also occurred in GWSS populations from the Carolinas and Texas.« less
  • Immunogenicity of Plasmodium gallinaceum sporozoites for chicks and their in vitro reactivity with normal and specific immune sera were studied. Two sporozoite populations recovered from experimentally infected Aedes fluviatilis were used: sporozoites from salivary glands and sporozoites from midgut oocysts. Populations seven to nine days old of sporozoites recovered from salivary glands were infective for all chicks until the chicks were three weeks old; however, sporozoites recovered from midguts containing oocysts infected these chicks only if isolated on days 8-9, but not on day 7 after the mosquitoes' infective blood meal. Infectivity of the sporozoites was lost after exposure tomore » ultraviolet (UV) light (30 min) or X-rays (13 krad). Inactivated sporozoites from both sources proved highly immunogenic to chicks that were immunized by several intravenous or intramuscular injections. These parasites elicited a strong humoral immune response in the chicks, as measured by the circumsporozoite precipitation (CSP) reaction. The levels of the CSP antibodies were similar with sporozoites from both sources, there being no detectable differences in the percentage of reactive sporozoites or the intensity of the CSP reaction with sera containing antibodies to either sporozoites from salivary glands or sporozoites from oocysts. These results provide the first evidence that avian malaria sporozoites express the circumsporozoite protein that has been extensively characterized in mammalian malaria (rodent, simian, human sporozoites). Furthermore, we observed that the yields of sporozoites obtained from mosquito midguts, on days 8 and 9 of the P. gallinaceum infection, were at least twice as great as those obtained by salivary gland dissection, even 20 days after a blood meal.« less