Partial characterization of Prunus Necrotic Ringspot Virus on apple in EGYPT

A severe isolate of Prunus necrotic ringspot virus (PNRSV) was isolated from apple orchards in the vicinity of Nubaria city, Beheira governorate, Egypt. Infected-apple trees showed chlorotic, necrotic ringspots, and shoot holes on leaves. Severely infected- trees withered, became useless, and were removed causing severe economic losses. Reverse transcriptase (RT) polymerase chain reaction (PCR), RT-PCR, using degenerate primer pair for the coat protein (CP) gene of Ilarvirus amplified products similar to those produced from peach and apricot isolates of PNRSV-infecting stone fruits). Dot blotting immuno-binding assay (DBIA showed a positive reaction between PNRSV-infected apple sap and an Egyptian antiserum for PNRSV. Purified preparation from infected leaves, using the electro-elution technique yielded nucleoprotein, which had Amax and Amin at 260 and 240 nm respectively. Electron microscopy examination showed spherical virions with ca. 26 nm in diameter.

Genetic characterization of a mild isolate of papaya ringspot virus type-P (PRSV-P) and assessment of its cross-protection potential under greenhouse and field conditions

A mild isolate of Papaya ringspot virus type-P, abbreviated as PRSV-mild, from Ecuador was sequenced and characterized. The most distinguishing symptom induced by PRSV-mild was gray powder-like leaf patches radiating from secondary veins. In greenhouse experiments, PRSV-mild did not confer durable protection against a severe isolate of the virus (PRSV-sev), obtained from the same field. Furthermore, isolate specific detection in mixed-infected plants showed that PRSV-sev becomes dominant in infections, rendering PRSV-mild undetectable at 90–120 days post superinfection. Virus testing using isolate-specific primers detected PRSV-mild in two out of five surveyed provinces, with 10% and 48% of incidence in Santo Domingo and Los Ríos, respectively. Comparative genomics showed that PRSV-mild lacks two amino acids from the coat protein region, whereas amino acid determinants for asymptomatic phenotypes were not identified. Recombination events were not predicted in the genomes of the Ecuadorean isolates. Phylogenetic analyses placed both PRSV-mild and PRSV-sev in a clade that includes an additional PRSV isolate from Ecuador and others from South America.

Inheritance of Resistance to Zucchini Yellow Mosaic Virus in Watermelon

Sources of resistance to the Zucchini yellow mosaic virus-Florida strain (ZYMV-FL) have been identified within the Citrullus genus. Inheritance of resistance to ZYMV-FL was studied in PI 595203 (Citrullus mucosospermus), a resistant watermelon accession. The F1, F2, and BC1 progenies derived from the cross ‘Calhoun Gray’ (CHG) × PI 595203 and ‘New Hampshire Midget’ (NHM) × PI 595203 were used to study the inheritance of resistance to ZYMV-FL. Seedlings were inoculated with a severe isolate of ZYMV-FL at the first true leaf stage and rated weekly for at least 6 weeks on a scale of 1 to 9 on the basis of severity of viral symptoms. A single recessive gene (zym-FL) was found to control the high level of resistance to ZYMV-FL in PI 595203.

Genomic of the severe isolate of Maize chlorotic dwarf virus

The complete sequences of the severe isolates of Maize chlorotic dwarf virus (MCDV-S) were obtained from cDNA clones of the viral RNA. The genome is 11 785 nucleotides (nt) in length and contains one large open reading frame between nt 455 and 10 825 that encodes a polyprotein of 3458 amino acids. Computer analysis revealed several conserved protein motifs similar to other plant picorna-like viruses. Comparison of the genome sequence of MCDV-S with that of the type strain of MCDV showed high similarities despite the difference in symptoms.  

Segregation of Distinct Variants from Citrus tristeza virus Isolate SY568 Using Aphid Transmission

A well-studied severe isolate of Citrus tristeza virus (CTV) known as SY568 has previously been shown to contain multiple variants of the virus which differ in their genetic and biological characters. Aphid transmission was used in an attempt to segregate some of these variants for further characterization. Resulting infections gave symptoms which varied from asymptomatic to more severe than the inoculum source. RNase protection assays (RPAs) were used to compare nine regions of the CTV genome and determine whether unique strains could be identified. Five aphid-transmitted subcultures, with fingerprints that were different from those of the inoculum sources in at least one genomic area, were then cloned, sequenced, and compared with known isolates. An asymptomatic strain was shown to be different in every area of the CTV genome when examined by RPA and sequencing of selected regions. Mixed-infection studies using graft transmission of the asymptomatic subculture and two of the more severe aphid-transmitted subcultures showed that the mild strain was not able to compete well when in the presence of any of the severe variants tested, and its titer was significantly reduced from that seen in single infection. The mild strain and a selected severe strain were singly graft inoculated into five different citrus hosts (sweet orange, grapefruit, sour orange, lemon, and lime), where they maintained their distinct biological and genetic characteristics.