First Report of Narcissus mosaic virus Infecting Crocus spp. Cultivars in the Netherlands

Abstract

A survey to identify virus diseases affecting Crocus spp. in the Netherlands was conducted during April 2004. Crocus spp. (cvs. Flavus, Pick-wick, Remembrance, and Grand Maitre) with symptoms suggestive of virus infection (stunting, yellowing, necrosis, and flower color breaking) were collected from several fields in the Breezand and Lisse districts in northern and southern Netherlands, respectively. All samples were tested for the presence of six known crocus-infecting viruses (1,2) using enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) assays. The ELISA assay was performed with the following polyclonal and monoclonal antibodies: Iris severe mosaic virus (ISMV); Tobacco rattle virus (TRV) isolates F, Y, and J obtained from the Applied Plant Research Institute, Lisse, Netherlands; Arabis mosaic virus; Cucumber mosaic virus from the Plant Research International Institute, Wageningen, Netherlands; Iris yellow spot virus (IYSV) from the Virology Department at Wageningen University, Netherlands; and the potyvirus group-specific monoclonal antiserum from the DSMZ, Braunschweig, Germany. All samples that tested positive with a potyvirus antiserum were further tested for the presence of Bean yellow mosaic virus (BYMV) using a BYMV-specific antiserum. Serological results obtained indicated that BYMV, detected with the potyvirus antiserum and BYMV-specific antiserum, and ISMV were the most commonly encountered viruses. Tobacco necrosis virus (TNV) and TRV were only found occasionally, whereas IYSV, was not detected in any of the samples tested. To study the presence of viruses not yet reported, total RNA was extracted and tested with a RT-PCR assay with carlavirus, potexvirus, necrovirus (R. Miglino, unpublished), and potyvirus (3) genus-specific oligonucleotides. In accordance with the ELISA results, PCR amplicons were obtained with the potyvirus, TNV, and TRV primer sets. Furthermore, a 280-bp amplicon corresponding to the expected size was amplified in a RT-PCR assay performed on total RNA with a potexvirus genus-specific primer set. The reverse primer (5′-AGC ATG GCG CCA TCT TGT GAC TG-3′) was located upstream in the conserved viral replicaseencoding region at position 4254-4231 of Narcissus mosaic virus (NMV) RNA genome (Genbank Accession No. D13747) and the forward primer (5′-CTG AAG TCA CAA TGG GTG AAG AA-3′) was located downstream at position 3969–3992. Sequence homology using BLAST analysis of the cloned and sequenced PCR product showed 98% identity with NMV. Although the virus has a very narrow host range, the results of this study may have a significant impact on the crocus industry in the Netherlands. To our knowledge, this is the first report of NMV infecting crocus. References: (1) M. G. Bellardi and A. Pisi. Inf. Fitopatol. 37:33, 1987. (2) A. F. L. M. Derks. Crocus spp. Pages 260–264 in: Virus and Virus-like Diseases of Bulbs and Flower Crops. G. Loebenstein et al., eds. Wiley publishers, West Sussex, UK, 1995. (3) S. A. Langeveld et al. J. Gen. Virol. 72:1531, 1991.

Transcriptome from saffron (Crocus sativus) plants in Jammu and Kashmir reveals abundant soybean mosaic virus transcripts and several putative pathogen bacterial and fungal genera

“…For example, tobacco MV is a Tobamovirus, while all other MVs in Table 1 are Potyviruses. Previously, saffron has been shown to be infected with bean yellow MV [18] and Narcissus MV [19]. Iris, which belong to the same Iridaceae family as saffron, was shown to be infected with Iris severe MV [20,21] and Narcissus MV [22].…”

Detection and molecular characterization of the Iris severe mosaic virus-Ir isolate from Iran

“…(Miglino et al 2005). The presence of potyvirus latent infections in C. sativus (Russo et al 1979) and C. cartwrightianus, and probably progenitor species of saffron (Caiola and Canini 2010), demonstrate an ancient infection of Crocus spp.…”

Narcissus yellow stripe virus and Narcissus mosaic virus detection in Narcissus via multiplex TaqMan-based reverse transcription-PCR assay

“…Visible viral symptoms of NMV include severe yellowing mosaic, leaf distortion and chlorosis, and flower colour breaking with unapparent symptoms during the early season (Miglino et al . ; He ; Hunter et al . ; Wylie et al .…”

“…NYSV-infected narcissus plants develop yellowing stunted stripes or mottle symptoms on the leaves as well as roughening, misshaping and possible flower streaking (Brunt and Atkey 1967;Chen et al 2007). Visible viral symptoms of NMV include severe yellowing mosaic, leaf distortion and chlorosis, and flower colour breaking with unapparent symptoms during the early season (Miglino et al 2005;He 2009;Hunter et al 2011;Wylie et al 2014a). Therefore, it proved difficult to distinguish NYSV and NMV only based on similar symptoms.…”