Development of a one-step RT-qPCR assay for the detection of Grapevine leafroll-associated virus 7.

Grapevine leafroll disease (GLD) is one of the most economically important viral diseases of grapevines. GLD is caused by a complex of several ssRNA (+) viruses referred to as Grapevine leafroll-associated viruses (GLRaVs). To date, five different GLRaV species have been identified. One of those species, GLRaV-7, was first reported from a symptomless white-fruited wine grape cultivar from Albania. Since its discovery, GLRaV-7 has been reported from 14 countries. Although serological assays have been developed to detect GLRaV-7, commercially available antibodies produce high background signals making them unsuitable for regulatory testing. Furthermore, while molecular detection assays have been shown to be more sensitive when compared to the serological assays, published molecular assays, except the one Reverse Transcription-quantitaive Polymerase Chain Reaction (RT-qPCR) assay based on heat shock protein 70 homologue (HSP70h) gene, have been reported to be inadequate in detecting all reported isolates of GLRaV-7. Availability of multiple assays provides flexibility to diagnostic laboratories in cases where the chosen assay fails to detect a strain or an isolate of a pathogen due to variation in its targeted region or where additional confirmation of the results is required. In this study, we developed a sensitive and specific RT-qPCR assay, based on a region of p61 gene of GLRaV-7, which detected all available isolates.

First Report of Hemp Canker Caused by Sclerotinia sclerotiorum in Alberta, Canada.

Hemp (Cannabis sativa L.) is a herbaceous annual grown mainly for its blast fiber and seed oil. In 1999, Health Canada issued licenses to plant 12,145 ha of hemp in Canada. Of these, 730 ha were in Alberta. During the last week of August, hemp plants (cv. Fasamo) in a central Alberta field showed the following symptoms and signs: wilting foliage turning light brown; dry tan to gray lesions on stems; shredding and breaking of stems at the lesion; presence of white mycelium in the lesion; and black round, irregular, or oblong sclerotia (up to 5 mm diameter and 2 to 11 mm long) present externally at the lesion on the stem and inside the pith cavity. Lesions were found at the crown, near the inflorescence, and along the entire stem length. Disease incidence in a survey of six commercial fields (40 ha) ranged from 1 to 8%. The organism isolated from lesions on potato dextrose agar produced white aerial mycelia and large numbers of sclerotia characteristic of Sclerotinia sclerotiorum. Pathogenicity was confirmed by inoculating 23-day-old greenhouse-grown hemp plants (cv. Fasamo) with autoclaved wheat grains colonized for 14 days with a S. sclerotiorum culture previously isolated from an infected hemp plant. The grains were placed on soilless growing medium near the plant and covered very lightly. One week after inoculation, grayish lesions appeared on the stems, white mycelia appeared on lesions, and plants wilted. The pathogen was reisolated from the lesions. This is the first report of S. sclerotiorum on hemp in Alberta, Canada. The disease known as hemp canker has been reported to cause severe losses under cool wet conditions in the Netherlands (1). Reference: (1) J. M. McPartland. J. Int. Hemp Assoc. 3:19, 1996.