scholarly journals Maintenance of coat protein N-terminal net charge and not primary sequence is essential for zucchini yellow mosaic virus systemic infectivity

2004 ◽  
Vol 85 (11) ◽  
pp. 3421-3430 ◽  
Author(s):  
Boaz Kimalov ◽  
Amit Gal-On ◽  
Ran Stav ◽  
Eduard Belausov ◽  
Tzahi Arazi
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Cell Movement ◽  
Yellow Mosaic Virus ◽  
Virus Infectivity ◽  
Amino Acid Residues ◽  
Primary Sequence ◽  
Net Charge ◽  
Truncation Mutant

Zucchini yellow mosaic virus (ZYMV) surface exposed coat protein (CP) N-terminal domain (Nt) is 43 aa long and contains an equal number of positively and negatively charged amino acid residues (CP-Nt net charge=0). A ZYMV-AGII truncation mutant lacking the first 20 aa of its CP-Nt (AGII-CPΔ20; CP-Nt net charge=+2) was found to be systemically non-infectious even though AGII mutants harbouring larger CP-Nt deletions were previously demonstrated to be fully infectious. Nevertheless, AGII-CPΔ20 infectivity was restored by fusion to its CP-Nt two Asp residues or a negatively charged Myc peptide, both predicted to neutralize CP-Nt net positive charge. To evaluate further the significance of CP-Nt net charge for AGII infectivity, a series of CP-Nt net charge mutants was generated and analysed for systemic infectivity of squash plants. AGII-CPKKK harbouring a CP-Nt amino fusion of three Lys residues (CP-Nt net charge=+3) was not systemically infectious. Addition of up to four Asp residues to CP-Nt did not abolish virus infectivity, although certain mutants were genetically unstable and had delayed infectivity. Addition of five negatively charged residues abolished infectivity (AGII-CPDDDDD; CP-Nt net charge=−5) even though a recombinant CPDDDDD could assemble into potyviral-like particle in bacteria. Neutralization of CP-Nt net charge by fusing Asp or Lys residues recovered infectivity of AGII-CPKKK and AGII-CPDDDDD. GFP-tagging of these mutants has demonstrated that both viruses have defective cell-to-cell movement. Together, these findings suggest that maintenance of CP-Nt net charge and not primary sequence is essential for ZYMV infectivity.

Field Evaluation of Transgenic Squash Containing Single or Multiple Virus Coat Protein Gene Constructs for Resistance to Cucumber Mosaic Virus, Watermelon Mosaic Virus 2 and Zucchini Yellow Mosaic Virus

1995 ◽  
Vol 13 (12) ◽  
pp. 1458-1465 ◽  
Author(s):  
David M. Tricoll ◽  
Kim J. Carney ◽  
Paul F. Russell ◽  
J. Russell McMaster ◽  
David W. Groff ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Protein Gene ◽  
Zucchini Yellow Mosaic Virus ◽  
Field Evaluation ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Multiple Virus ◽  
Transgenic Squash ◽  
Virus Coat

scholarly journals Isolation and Molecular Characterization of a Chilean Isolate of Zucchini yellow mosaic virus

Plant Disease ◽  
2001 ◽  
Vol 85 (6) ◽  
pp. 644-648 ◽  
Author(s):  
H. Prieto ◽  
A. Bruna ◽  
P. Hinrichsen ◽  
C. Muñoz
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sequence Comparisons ◽  
Severe Stunting ◽  
Chilean Isolate

Zucchini yellow mosaic virus (ZYMV) was described in 1981 affecting squash, melon, and other cultivated cucurbits with severe stunting and yellowing symptoms. It was reported to be present in most countries where cucurbits are grown, and in Chile since 1995, from surveys using enzyme-linked immunosorbent assay (ELISA) but without further characterization. A potyvirus was isolated from ELISA-positive symptomatic plants. The results indicate that this virus is ZYMV based on symptoms on herbaceous indicators, immunospecific electron microscopy of the purified particle, and sequencing of 395 bases of the 3′ end of the coat protein gene. The virus was detected in melon, watermelon, and squash plants. In agreement with previous descriptions for ZYMV, the Chilean isolate is a flexuous filamentous particle 740 nm long with one main protein of approximately 36 kDa. Nucleotide sequence comparisons of the 3′ portion of the coat protein gene revealed a high similarity to the Connecticut and California strains.

scholarly journals First Report of Zucchini yellow mosaic virus Associated with Leaf Crinkle and Yellow Mosaic Diseases of Cucurbit Plants in Mali

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 923-923 ◽  
Author(s):  
W. S. Tsai ◽  
I. K. Abdourhamane ◽  
D. Knierim ◽  
J. T. Wang ◽  
L. Kenyon
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
The United States ◽  
Yellow Mosaic Virus ◽  
Nucleotide Identity ◽  
Watermelon Mosaic Virus ◽  
Virus Species ◽  
First Report

The aphid-transmitted Zucchini yellow mosaic virus (ZYMV; genus Potyvirus, family Potyviridae) has been reported to cause severe epidemics and yield losses in cucurbit crops worldwide (1). In Africa, ZYMV has been detected in Algeria, Egypt, Madagascar, Mauritius, Mayotte, Morocco, Nigeria, Reunion, South Africa, Sudan, Swaziland, and Tunisia (1). In April 2009, leaf yellowing, mosaic, crinkling, and curling were common on cucurbit plants in fields in Mali. Symptomatic leaf samples were collected from five cucumber (Cucumis sativus) plants in Kati, two watermelon (Citrullus lanatus) plants in Samanko, and one weedy melon (Cucumis sp.) plant in Baguineda. All samples tested positive for ZYMV and were negative for Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV), Papaya ringspot virus type W (PRSV-W), Watermelon mosaic virus (WMV), and Watermelon silver mottle virus (WSMoV) by double-antibody sandwich (DAS)-ELISA. They also tested negative for Melon yellow spot virus (MYSV) by indirect ELISA. Antibodies against ZYMV and WMV were obtained from DSMZ, Braunschweig, Germany, and those against CGMMV, MYSV, PRSV-W, and WSMoV were provided by Shyi-Dong Yeh, National Chung Hsing University, Taichung, Taiwan. Six ZYMV ELISA-positive samples (three cucumber, two watermelon, and the weedy melon sample) were also tested by reverse transcription (RT)-PCR using the potyvirus universal primer pair Sprimer1/Oligo(dT) (2). The expected 1.6-kb viral cDNA was amplified from all six samples and each was sequenced. All sequences obtained from cucumber (GenBank Accession Nos. HM005307, HM005308, and HM005309), watermelon (GenBank Accession Nos. HM005311 and HM005312), and weedy melon (GenBank Accession No. HM005310) isolates were 1,684 nucleotides (nt) long excluding the 3′ poly-A tails. They comprised the 3′-terminal of the NIb region (1 to 633 nt), the coat protein region (634 to 1473 nt), and the 3′-untranslated region (1,474 to 1,684 nt). Because the sequences shared high nucleotide identity (98.3 to 99.7%), these isolates were considered to be the same virus species. When the sequences were compared by BLASTn searching in GenBank and analyzed by DNAMAN Sequence Analysis Software (Lynnon Corporation, St-Louis, Pointe-Claire, Quebec, Canada), they were found to have the greatest nucleotide identity (97.4 to 98.0%) with the Connecticut strain of ZYMV (ZYMV-Connecticut; GenBank Accession No. D00692), within a clade of isolates from China, Italy, Japan, and the United States. When assessed separately, their coat protein (97.7 to 98.3% nucleotide and 98.9 to 99.6% amino acid identity) and 3′-untranslated regions (96.7 to 97.2% identity) also had greatest homology with ZYMV-Connecticut. To our knowledge, this is the first report of ZYMV infecting cucurbit plants in Mali. ZYMV should be taken into consideration when breeding cucurbit crops for this region, and managing viral diseases. References: (1) C. Desbiez et al. Plant Pathol. 46:809, 1997. (2) W. S. Tsai et al. Plant Dis. 94:378, 2010.

scholarly journals GENETIC ENGINEERING OF THE ZUCCHINI YELLOW MOSAIC VIRUS COAT PROTEIN GENE FOR EXPRESSION IN PLANTS.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1130d-1130
Author(s):  
Guowei Fang ◽  
Rebecca Grumet
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Zucchini Yellow Mosaic Virus ◽  
In Vitro Transcription ◽  
Yellow Mosaic Virus ◽  
35S Promoter ◽  
Plant Expression Vector

Zucchini yellow mosaic virus (ZYMV), a potyvirus, can cause major losses in cucurbit crops. With the goal of genetically engineering resistance to this disease we have engineered the ZYMV coat protein gene into a plant expression vector. The complete coat protein coding sequence, or the conserved core portion of the capsid gene, was attached to the 5' untranslated region of tobacco etch virus (TEV) in the pTL37 vector (Carrington et al., 1987, Nucl. Acid Res. 15:10066) The capsid constructs were successfully expressed by in vitro transcription and translation systems as verified by SDS-PAGE and ZYMV coat protein antibody. The constructs were then subcloned using polymerase chain reaction and attached to the CaMV 35 S transcriptional promoter on the CIBA-GEIGY pCIB710 plasmid. The constructs containing the CaMV 35S promoter, the 5' untranslated leader of TEV, and ZYMV coat protein sequences were then put between the Agrobacterium tumefaciens left and right borders in the pCIB10 vector and transferred to A. tumefaciens strain LBA4404 by triparental mating. These vectors are now being used to transform muskmelon and cucumber; resultant transgenic plants will be tested for ZYMV coat protein expression.

scholarly journals Fevillea trilobata as a Natural Host of Zucchini yellow mosaic virus in Brazil

Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1261-1261 ◽  
Author(s):  
L. S. Boiteux ◽  
D. M. A. Spadotti ◽  
J. A. M. Rezende ◽  
E. W. Kitajima
Keyword(s):  
Amino Acid ◽  
Coat Protein ◽  
Mosaic Virus ◽  
Direct Sequencing ◽  
Zucchini Yellow Mosaic Virus ◽  
Amino Acid Sequences ◽  
Natural Host ◽  
Yellow Mosaic Virus ◽  
Biodiesel Fuel ◽  
Thin Sections

The antidote vines or nhandirobas (Fevillea trilobata L. [Cucurbitaceae]) are dioecious plant species native to the South American Neotropics (1). Genetic materials of these species are now being domesticated and evaluated as potential crops for seed-oil extraction aiming to produce biodiesel fuel (2). Plants of F. trilobata (Accession No. CNPH-001) were cultivated from seeds under open field conditions during the years 2008 through 2011 in Brasília-DF, Brazil. Approximately 200 plants exhibiting mosaic symptoms and severe leaf malformation (with typical bubble-like patches) were found in all fields every year. Apical mosaic was slightly more severe in female than in male plants. Electron microscopy examination of negatively stained extracts of symptomatic leaf tissue showed the presence of filamentous particles about 700 to 800 nm long. Analysis of ultra-thin sections of the same tissues revealed the presence of lamellar inclusions typical of a potyvirus infection. No aphid colonies were observed on field-grown F. trilobata plants. The virus was mechanically transmitted to healthy Cucurbita pepo cv. Caserta and Luffa cylindrica, causing systemic mosaic. Sap from these infected plants reacted in PTA-ELISA with polyclonal antiserum against Zucchini yellow mosaic virus (ZYMV), but not with antisera against Papaya ringspot virus – type W (PRSV-W), Cucumber mosaic virus (CMV), and Zucchini lethal chlorosis virus (ZLCV). Total RNA extracted from experimentally infected C. pepo was analyzed by RT-PCR using specific pairs of primers for the coat protein gene of ZYMV (3). A cDNA fragment of approximately 1,186 bp was amplified and the nucleotide sequence obtained by direct sequencing. Comparisons of the nucleotide (837 nt) and deduced amino acid (279 aa) sequences of the coat protein genomic segment (GenBank Accession No. JX502677) revealed 93 to 98% and 97 to 98% identity, respectively, with the corresponding nucleotide and amino acid sequences of a group of ZYMV isolates from distinct hosts (AY188994, AY279000, and NC_003224). The infection by ZYMV might cause fruit yield losses to F. trilobata. In addition, the infected F. trilobata crops might work as a reservoir of ZYMV providing inoculum to other cucurbit hosts since it has been managed as a semi-perennial crop. To our knowledge, this is the first report of the genus Fevillea as a natural host of ZYMV. References: (1) M. Nee et al. Syst. Bot. 34:704, 2009. (2) E. G. Shay. Biomass Bioenergy 4:227, 1993. (3) K. G. Thomson et al. J. Virol. Meth. 55:83, 1995.

scholarly journals Cloning and characterisation of nanobodies against the coat protein of Zucchini yellow mosaic virus

2018 ◽  
Vol 54 (No. 4) ◽  
pp. 215-221 ◽  
Author(s):  
Zakri Adel M ◽  
AL-Doss Abdullah A ◽  
Sack Markus ◽  
Ali Ahmed A ◽  
Samara Emad M ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Viral Infections ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Diagnostic Tools ◽  
Diagnostic Assays ◽  
The Family ◽  
Variable Domains ◽  
Heavy Chain Antibody

Zucchini yellow mosaic virus (ZYMV), in the family Potyviridae, causes an economically important disease. Antibodies are valuable reagents for diagnostic assays to rapidly detect viral infection. Here, we report the isolation of camel-derived variable domains of the heavy chain antibody (VHH, also called nanobodies) directed against the coat protein (CP) of ZYMV. Several nanobodies that specifically recognise ZYMV-CP were identified. The isolated nanobodies showed binding not only to recombinant ZYMV-CP but also to native ZYMV, indicating that these nanobodies can be used in diagnostic tools to detect viral infections.

scholarly journals Rapid evolutionary dynamics of zucchini yellow mosaic virus

2008 ◽  
Vol 89 (4) ◽  
pp. 1081-1085 ◽  
Author(s):  
Heather E. Simmons ◽  
Edward C. Holmes ◽  
Andrew G. Stephenson
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Evolutionary Dynamics ◽  
Rna Viruses ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Nucleotide Substitutions ◽  
The Family ◽  
Coalescent Approach

Zucchini yellow mosaic virus (ZYMV) is an economically important virus of cucurbit crops. However, little is known about the rate at which this virus has evolved within members of the family Cucurbitaceae, or the timescale of its epidemiological history. Herein, we present the first analysis of the evolutionary dynamics of ZYMV. Using a Bayesian coalescent approach we show that the coat protein of ZYMV has evolved at a mean rate of 5.0×10−4 nucleotide substitutions per site, per year. Notably, this rate is equivalent to those observed in animal RNA viruses. Using the same approach we show that the lineages of ZYMV sampled here have an ancestry that dates back no more than 800 years, suggesting that human activities have played a central role in the dispersal of ZYMV. Finally, an analysis of phylogeographical structure provides strong evidence for the in situ evolution of ZYMV within individual countries.

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