Sequence analysis of RNA-2 of different isolates of broad bean wilt virus confirms the existence of two distinct species

Abstract

The complete nucleotide sequence of RNA-2 from a Japanese isolate IP of broad bean wilt virus (BBWV) was determined. The sequence encodes a single large polyprotein, which contains a putative movement protein and two coat proteins (CPs). The 3'-terminal sequences of RNA-2 were also determined for three other Japanese isolates and two ATCC isolates (PV132 and PV176) of BBWV. The CPs of the four Japanese isolates share 86.8-98.0% amino acid sequences homology with one another and 88.3-96.5% with those reported for the isolate PV131 (BBWV-2). However, they have only 57.9-66.2% homology with those of PV132 and PV176 (BBWV-1).

Virus Infections in Gentiana Species

“…Green and D.-E. Lesemann, unpublished) and Megakepasma from Singapore (S.M. Wong and D.-E. Lesemann, unpublished), and reports on BBWV2 in China (Xu et al, 1988) and Japan (Kobayashi et al, 1999) indicate that Asia is the continent of origin of this virus. On the other hand, the related BBWV1 is a common virus of wild plants and ornamentals in Germany, and was also recorded in Eustoma russellianum (syn.…”

“…It was mainly detected in vegetatively propagated plants of G. scabra. These in vitro propagated plants originate in Japan, where the virus was already isolated from gentian before (Nagao et al, 1978;Yamashita, 1990;Kobayashi, 1999). So far, in the Netherlands and in Germany BBWV2 was detected only in imported plants from Petunia originating in Japan and in Gentiana and Verbena .…”

Detection and identification of Fabavirus species by one-step RT-PCR and multiplex RT-PCR

“…The availability of nucleotide sequences of BBWV-1 (Kobayashi et al, 1999a(Kobayashi et al, ,b, 2003Ferrer et al, 2005), BBWV-2 (Ikegami et al, 1998(Ikegami et al, , 2000Nakamura et al, 1998;Kuroda et al, 2000;Lee et al, 2000;Qi et al, 2000a,b;Koh et al, 2001;Ferrer et al, 2011), GeMV (Kobayashi et al, 2005) and CuMMV (Dong et al, 2012) has enabled the development of more sensitive and specific diagnostic methods based on molecular hybridization and reverse transcription, followed by polymerase chain reaction (RT-PCR). These techniques include: (1) molecular hybridization for the detection of BBWV-1 (Ferrer et al, 2008); (2) real time RT-PCR for specific detection and quantification of BBWV-1 and BBWV-2 ; and (3) simultaneous detection and identification of BBWV-1, BBWV-2 and GeMV using conserved primers (Ferrer et al, 2007).…”

“…The primers for multiplex RT-PCR were designed based on: (1) nucleotide sequences specific to each virus, but conserved between isolates; (2) compatibility between primers so that they could be used in the same RT-PCR; and (3) differences in RT-PCR product size from each virus, so that they could be identified by electrophoresis. The design process to obtain specific primers for BBWV-1, BBWV-2 and GeMV was very complex due to the high genetic variability of these viruses (Kobayashi et al, 1999a(Kobayashi et al, ,b, 2005Ferrer et al, 2005Ferrer et al, , 2011Wang et al, 2008) in comparison with most other plant viruses (García-Arenal et al, 2001;Rangel et al, 2011;Tiberini et al, 2011;Davino et al, 2012). The primers were designed to ensure that the expected sizes of the RT-PCR products were approximately 622, 510, 370, 238 and 98 bp for LMMV, BBWV-1, GeMV, BBWV-2 and CuMMV, respectively (Table 2).…”

Caracterización molecular y desarrollo de métodos de diagnóstico del género Fabavirus Evaluación de BTH como método de control

“…Sin embargo, los estudios biológicos se realizaron hace mucho tiempo (entre 1947 y 1982), con muy pocos aislados y de manera poco sistemática (no se realizaron ensayos comparativos de varios aislados), por lo que no está claro si las diferencias biológicas observadas ocurrieron solamente en los aislados estudiados o si son atribuibles a especies distintas. Posteriormente, la determinación de la secuencia nucleotídica del genoma de algunos aislados confirmó que BBWV-1 y BBWV-2 son dos especies distintas (Kobayashi et al, 1999) pero todavía no se dispone de ninguna secuencia nucleotídica de LMMV. Recientemente, se ha descubierto un nuevo serotipo al que se ha propuesto como una nueva especie, Gentian mosaic virus (GeMV), basándose en su secuencia nucleotídica (Kobayashi et al, 2005).…”