Hervé Lapierre

Asymmetrical Distribution of Barley Yellow Dwarf Virus PAV Variants Between Host Plant Species

ABSTRACT A large epidemiological study of the genetic variation of barley yellow dwarf virus (BYDV) serotype PAV involving different host plant species was conducted. French BYDV PAV isolates were collected from barley and ryegrass, and their capsid protein gene sequences characterized using restriction fragment length polymorphism, single-strand conformation polymorphism, and sequence analyses. The data show that BYDV PAV isolates from five different continents are separated into two distinct groups named cpA and cpB, which are distributed irrespective of geographical location. Amino acid identity of the capsid proteins ranged from 93 to 99.5% in group cpA and from 95 to 99.5% in group cpB, while this value was only from 82 to 88% between the groups. Moreover, isolates from each group were found preferentially (up to 98%) in one of the two plant species examined. These results show that host plant species play a role in isolate selection and maintenance and that they contribute to the genetic diversity of BYDV PAV.

Comparison and differentiation of Wheat Yellow Mosaic Virus(WYMV), Wheat Spindle Streak Mosaic Virus (WSSMV) and Barley Yellow Mosaic Virus (BaYMV) isolates using WYMV monoclonal antibodies

Twelve monoclonal antibodies (MAbs) were obtained by immunizing mice with a French isolate (F1) of wheat yellow mosaic virus (WYMV). Three of these (3D12, 2C1, 6C3) belong to the IgM class and the nine others to the IgG class (3D8, 3H1, 2B8, 1F2, 3C10, 4F12, 3H9, 1G5, 54). In antigen-coated plate (ACP) ELISA and indirect double antibody sandwich (IDAS) ELISA, all MAbs recognize the WYMV (F1) both in the form of purified particles and in wheat leaf extract. The analysis of numerous French isolates of WYMV shows a variable reactivity with MAbs 3D8, 3H1, 2B8, 3C10, 3H9 and 1G5 in IDAS — and ACP-ELISA. The Japanese isolate of WYMV and United States isolates of wheat spindle streak mosaic virus (WSSMV) were detected in IDAS- and ACP-ELISA by ten of the MAbs tested showing that the wheat bymoviruses originating from the three locations share a high epitopic homology. French isolates of barley yellow mosaic virus (BaYMV; pathotypes 1 and 2) were only detected in ACP-ELISA with MAbs 6C3, 3D8, 3H1 and 2B8 whereas the two Japanese strains (I-1, II-1) of MaYMV were recognized with these and also with that of 3C10. In IDAS-ELISA, the two Japanese strains were clearly detected by MAbs, 6C3, 3D8, 3H1, 1F2, 3C10 and 1G5 and the British and Belgian (pathotype 2) isolates only by that of 6C3. Only the Japanese strain of BaYMV, 1-1 could be detected with MAb 3H9 in this ELISA system.

Aubian wheat mosaic virus, a new soil-borne wheat virus emerging in France

The properties of Aubian wheat mosaic virus (AWMV), a new soil-borne wheat virus in France, were investigated. Symptoms include foliar mosaic and severe stunting of winter wheat. The vector of the disease is unknown but the plants infected carry Polymyxa graminis in the roots. AWMV was transmitted mechanically to wheat and to two dicotyledoneous species: Lactuca sativa and Vicia faba. This virus was transmitted by seed to three winter wheat cultivars tested. Purified preparations contained rod-shaped particles with a variable length of 150–700 nm. Certain particles are very long and appear flexible. Antiserum raised against AWMV reacted specifically with AWMV in both indirect and direct enzyme-linked immunosorbent assays (ELISA). The incidence of AWMV in 26 winter wheat cultivars was investigated in the field during the growing season of 1999–2000. AWMV was detected in roots and shoots of all cultivars regardless of the symptoms. Twelve virus species belonging to the genera Benyvirus, Bymovirus, Furovirus, Pecluvirus and Pomovirus did not react with the AWMV antisera. A new tubular virus described in winter wheat in Bedfordshire in England reacted strongly with AWMV in ELISA. It is concluded that AWMV and probably the Bedford-virus constitute a previously undescribed tubular virus biologically and serologically distinct from other soil-borne viruses of wheat.

Sequences of the 3′ Halves of the Genomes of Barley Yellow Dwarf Virus-PAV cpA Isolates that Vary in Symptom Severity

Barley yellow dwarf virus (BYDV)-PAV isolates from USA have been separated into two distinct clusters (Chay et al. (1996) Virology 219: 57–65; Chay et al. (1996) Phytopathology 86: 370–377). Following this finding we have shown that BYDV-PAV is divided into two groups cpA and cpB based on their coat protein gene sequence, and distinct host preferences (Mastari et al. (1998) Phytopathology 88: 818–821). We have sequenced the complete 3′ half of the genomes of two lethal and two mild cpA isolates and compared them with those of several known PAV cpA isolates to assess variability and locate potential determinants of severity. Open reading frames (ORFs) 3, 4, 5, 6 and the 3′ untranslated regions had different percent homologies between isolates: ORF5 (92–97%), ORF3 (88–98%) 3′-translational enhancer (87–100%) ORF4 (85–99%), 3′ untranslated region (72–97%) and ORF6 (61–99%). In contrast to the mild isolates, the field-lethal isolates (FHv1 and FHv2) fell into the same cluster, regardless of the genomic region analysed. The isolates FHv1 and FHv2 differed from mild isolates by eight amino acid substitutions in ORFs 3 and 4, and insertions in ORF5. Four amino acid substitutions in the 17-kDa protein encoded by ORF4 caused a change in local net charge in the field-lethal isolates. Two insertions of four amino acids were identified in the C-terminal half of ORF5 of the field-lethal isolates, but were not present systematically in all lethal isolates analysed. The potential relationships of these differences in predicted amino acid sequences to disease severity are discussed.