The main aims of this study were to determine the overwintering form and the primary inoculum of Erysiphe necator, the causal agent of grapevine powdery mildew in southern Syria. Eleven vineyards located at five different geographical sites were visited every week from March to November in 2014 and 2015. The results of field and histopathological studies showed that E. necator survived as mycelium in dormant grapevine buds during the winter season. The first flag shoots were observed shortly after bud break in spring. The number of flag shoots varied greatly according to vineyards, cultivars and years, being present on 27.4 to 61.9% of the grapevines in 2014, and on 5.2 to 40% of the grapevines in 2015. The percentage of flag shoots on the same grapevine also varied according to the year, cultivar and location. It was between 4.3 to 9.4% in 2014, and 2.1 to 3.6% in 2015. The disease was observed only on Balady and Black cultivars. Conidia were released from the second week of May to early September. The first conidia were trapped around mid-May, and the first secondary symptoms were observed on leaves from mid-May to early June according to the site. Chasmothecia were observed on leaves in 45.5% of the studied vineyards. The first observation of chasmothecia on leaves was in July, and their numbers varied greatly between vineyards and years. Chasmothecia were not detected on bark, nor were ascospores trapped at the beginning of the season. These results confirmed that the ascospores did not have any role in the initiation of spring infection. To the best of our knowledge, this is the first report of the overwintering form of E. necator in Syria