Brigitte Delecolle

Effects of onion yellow dwarf and leek yellow stripe viruses on symptomatology and yield loss of three french garlic cultivars

This study was conducted to determine the effect of two potyviruses, onion yellow dwarf virus (OYDV) and leek yellow stripe virus (LYSV), on the symptoms, growth, and potential yield loss of garlic (Allium sativum). For 2 consecutive years, the impact on leaf length, pseudostem diameter, and bulb weight was evaluated after mechanical inoculation of cultivars Messidrome, Germidour, and Printanor, the three main garlic cultivars grown in France. The reduction in bulb weight due to OYDV ranged from 39% for Germidour to about 60% for the two other cultivars. For LYSV, the reduction in bulb weight was less on Messidrome (17%) and Germidour (26%) than on Printanor (54%). Coinfection with both viruses further reduced growth and bulb weight. When cloves originating from bulbs infected by each virus alone or a mixture of both viruses were planted, results indicated that such chronic infection induced further yield reduction. An assay designed to evaluate the role of LYSV inoculation date on yield revealed that yield losses were the lowest for late-season infections. However, yield loss was greater than 30% when the inoculation was performed at the end of April, the time when natural contamination generally occurs in southern France. A comparison of the impact of mixed infections of OYDV and LYSV from different origins suggested that the isolates did not differ significantly in their effects on yield loss.

Cytological and molecular evidence that the whitefly-transmitted Cucumber vein yellowing virus is a tentative member of the family Potyviridae

Cucumber vein yellowing virus (CVYV) is widespread in cucurbits in the Middle East. CVYV has filamentous particles and is transmitted by Bemisia tabaci by the semi-persistent mode. It has not yet been assigned to a specific genus or family. Ultramicroscopic observations revealed numerous cylindrical cytoplasmic inclusions in melon and cucumber cells infected by CVYV isolates from Israel and Jordan. Depending on the section orientation, the inclusions appeared as pinwheels or as bundles. In addition, a 1.9 kb DNA fragment was amplified by RT-PCR from CVYV-infected plant extracts using primers designed to detect all potyvirids. Sequence comparisons with the amplified fragment indicated that CVYV is more closely related to Sweet potato mild mottle virus than to any other virus in the family Potyviridae: These results suggest that CVYV can be considered as a tentative new member of the genus Ipomovirus:, family Potyviridae:

Interaction between potyvirus helper component-proteinase and capsid protein in infected plants.

Monoclonal antibodies were raised against helper component-proteinase (HcPro) purified from plants infected with the potyvirus Lettuce mosaic virus (LMV). These antibodies were used in a two-site triple antibody sandwich ELISA assay together with polyclonal antibodies directed against purified virions. An interaction between HcPro and the viral coat protein (CP) was demonstrated in extracts of LMV-infected leaves, as well as for two other potyviruses, Plum pox virus and Potato virus Y. The CP-HcPro interaction was not abolished in LMV derivatives with an HcPro GFP N-terminal fusion, or with a deletion from the CP of the amino acids involved in aphid transmission. Electron microscopy indicated that HcPro probably does not interact with the CP in the form of assembled virions or virus-like particles. Together, these results suggest that the interaction detected between CP and HcPro might be involved in a process of the potyvirus cycle different from aphid transmission.

Use of modified plum pox virus coat protein genes developed to limit heteroencapsidation-associated risks in transgenic plants.

Aphid transmission of a non-aphid-transmissible strain of zucchini yellow mosaic virus (ZYMV-NAT) occurs in transgenic plants expressing the plum pox potyvirus (PPV) coat protein (CP) gene. Heteroencapsidation has been shown to be responsible for this modification in the epidemiological characteristics of the infecting virus. In order to prevent this biological risk, several modified PPV CP constructs were produced that were designed to interfere with heteroencapsidation itself or to block aphid transmission of heteroencapsidated virions. These constructs were first expressed in Escherichia coli in order to check for the accumulation of pseudo-particles by electron microscopy. Virus-like particles (VLPs) were found with the full-length CP and with a PPV CP lacking the DAG amino acid triplet involved in aphid transmission. However, no VLPs were observed with CP lacking R220, Q221 or D264, amino acids known to be essential for the assembly of other potyvirus CPs. Transgenic Nicotiana benthamiana lines expressing the different PPV CP constructs were infected with ZYMV-NAT. Aphid transmission assays performed with these plants demonstrated that the strategies developed here provide an effective means of minimizing the biological risks associated with heteroencapsidation.

Biological and Molecular Characterization of Moroccan watermelon mosaic virus and a Potyvirus Isolate from Eastern Sudan

A potyvirus (Su-94-54) was isolated from a naturally infected snake cucumber (Cucumis melo var. flexuosus) plant with severe mosaic and leaf deformation symptoms collected in Eastern Sudan. This isolate has a host range limited to cucurbits and is serologically distantly related to Moroccan watermelon mosaic virus (MWMV) and to Papaya ringspot virus (PRSV). Coat protein sequence analysis of Su-94-54 and MWMV and comparison with other potyviruses indicate that Su-94-54 is more closely related to MWMV than to any other potyvirus. Based on the amino acid sequence identity in the core part of the coat protein with MWMV (86%), this isolate could be regarded as a distinct species. However, because of biological, cytological, and serological affinities with MWMV, we propose that this isolate be considered as a strain of MWMV, possibly an evolutionary intermediate between MWMV and PRSV, until more is known on the structure of the PRSV subgroup within the genus Potyvirus.

Characterization, nucleotide sequence and genome organization of leek white stripe virus, a putative new species of the genus Necrovirus.

SummaryWhite stripe is a disease affecting leek in France with which an isometric virusc. 30 nm in diameter is associated. The most evident symptom is the presence of white stripes on the leaves extending to the stem. Attempts to demonstrate transmission through the soil by sowing or transplanting leek in contaminated soil were unsuccessful. The virus was transmitted by sap inoculation to a narrow range of herbaceous hosts, all of which were infected only locally. Virus purification was from infected leek tissues, where it accumulated in large amounts, as demonstrated by ultrastructural observations. RNA was extracted from purified virus preparations and cDNA clones were prepared. The complete nucleotide sequence of the viral RNA was determined: The genome is 3662 nucleotides long and contains five open reading frames (ORFs). The first (ORF 1) encodes a putative translation product of Mr 23803 (p24) and read through of its amber stop codon results in a protein of Mr 82625 (p83) (ORF 2). ORF 3 and ORF 4 encode two small polypeptides of Mr 11280 (p11) and Mr 6261 (p6), respectively. ORF 5 encodes the capsid protein of Mr 27460 (p27). The genome organization and sequence alignments with the corresponding products of necroviruses suggest that the virus isolated from leek is a new species in the genusNecrovirus, for which the name of leek white stripe virus (LWSV) is proposed.

Melon Rugose Mosaic Virus: Characterization of an Isolate from Sudan and Seed Transmission in Melon

Melon rugose mosaic virus (MRMV) was isolated from snake cucumber (Cucumis melo var. flexuosus) in the Kassala region of Sudan in 1993. The host range of the virus was mostly limited to cucurbits, where it induced severe mosaic and leaf deformations. Cytopathological studies revealed severe chloroplast alterations, including vesicles at their periphery and the tendency to aggregate, which are typical of tymovirus infections, providing further evidence that MRMV is a tentative member of the genus Tymovirus. In melon and snake cucumber, MRMV was found to be seed transmitted at rates of 0.9 and 3.8%, respectively. Seed dissection experiments revealed that the virus could be detected in the seed coat, papery layer, and embryo. Seed disinfection treatments did not reduce seed transmission rates, which suggests an internal transmission. A preliminary screening for resistance in melon revealed some resistance in two out of 367 accessions tested.

Snake melon asteroid mosaic virus, a Tentative New Member of the Genus Sobemovirus Infecting Cucurbits

A virus isolate (Su-95-67) was obtained from a snake melon (Cucumis melo var. flexuosus) plant presenting severe chlorotic spots, mosaic, stunting, and leaf deformations collected in Eastern Sudan in 1995. Su-95-67 was easily mechanically transmissible and had a host range limited to a few cucurbit species. Isometric virus particles approximately 30 nm in diameter were observed in leaf dip preparations. A cytopathological study did not reveal alterations specific for a virus genus or family. A polyclonal antiserum was obtained and used in double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Su-95-67 was transmitted by seed at a low rate, by the red melon beetle (Aulacophora foveicollis), but not by the melon aphid (Aphis gossypii). Because Su-95-67 shared several properties with sobemoviruses, generic Sobemovirus reverse-transcription polymerase chain reaction primers were developed. They allowed amplification of a 384-bp fragment from extracts of plants infected by two sobemoviruses or by Su-95-67 but not from healthy plants extracts. Sequence comparison confirmed that Su-95-67 belongs to a new tentative Sobemovirus species for which we propose the name Snake melon asteroid mosaic virus (SMAMV). DAS-ELISA tests conducted on extracts of virus-infected cucurbit plants collected from 1992 to 2003 revealed the presence of SMAMV in 10.2% of 600 samples originating from different regions of Sudan.